Pharmaceutical compositions of 7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino [2,3-b]pyrazin-2(1h)-one, a solid form thereof and methods of their use

ABSTRACT

Provided herein are compositions of 7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one, solid forms, isotopologues and metabolites thereof, and methods of their use for the treatment of a disease, disorder, or condition.

This application is a divisional of U.S. application Ser. No.15/708,222, filed Sep. 19, 2017, currently allowed, which is adivisional of U.S. application Ser. No. 15/427,242, filed Feb. 8, 2017,now U.S. Pat. No. 9,795,603, issued Oct. 24, 2017, which is a divisionalof U.S. application Ser. No. 14/288,521, filed May 28, 2014, now U.S.Pat. No. 9,604,939, issued Mar. 28, 2017, which claims the benefit ofU.S. Provisional Application No. 61/828,506, filed May 29, 2013, theentire contents of which are incorporated herein by reference.

1. FIELD

Provided herein are compositions of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,a solid form thereof, isotopologues thereof, and methods of their usefor the treatment of a disease, disorder, or condition.

2. BACKGROUND

The connection between abnormal protein phosphorylation and the cause orconsequence of diseases has been known for over 20 years. Accordingly,protein kinases have become a very important group of drug targets. SeeCohen, Nature, 1:309-315 (2002). Various protein kinase inhibitors havebeen used clinically in treating a wide variety of diseases, such ascancer, chronic inflammatory diseases, diabetes, and stroke. See Cohen,Eur. J. Biochem., 268:5001-5010 (2001), Protein Kinase Inhibitors forthe Treatment of Disease: The Promise and the Problems, Handbook ofExperimental Pharmacology, Springer Berlin Heidelberg, 167 (2005).

The elucidation of the intricacy of protein kinase pathways and thecomplexity of the relationship and interaction among and between thevarious protein kinases and kinase pathways highlights the importance ofdeveloping pharmaceutical agents capable of acting as protein kinasemodulators, regulators, or inhibitors that have beneficial activity onmultiple kinases or multiple kinase pathways. Accordingly, there remainsa need for new kinase modulators.

The protein named mTOR (mammalian target of rapamycin), also known asFRAP, RAFTI or RAPT 1, is a Ser/Thr protein kinase related to the lipidkinases of the phosphoinositide 3-kinase (PI3K) family. It functions asa sensor of mitogen, energy, and nutrient levels; and is a centralcontroller of cell growth. mTOR has been shown to be one of the mostcritical proteins in the mTOR/PI3K/Akt pathway that regulates cellgrowth and proliferation. Georgakis and Younes, Expert Rev. AnticancerTher. 6(1):131-140 (2006). mTOR exists in two complexes, mammaliantarget of rapamycin complex 1 (mTORC1) which complexes with raptor, andmammalian target of rapamycin complex 2 (mTORC2) which complexes withrictor. While mTORC1 is sensitive to rapamycin analogs (such astemsirolimus or everolimus), mTORC2 is largely rapamycin-insensitive(Kim et al., Cell 110(2):163-175 (2002); Sarbassov et al., Science307:1098-1101 (2005)).

Several mTOR inhibitors have been or are being evaluated in clinicaltrials for the treatment of cancer. For example, temsirolimus wasapproved for use in renal cell carcinoma in 2007 and sirolimus wasapproved in 1999 for the prophylaxis of renal transplant rejection.Everolimus was approved in 2009 for renal cell carcinoma patients thathave progressed on vascular endothelial growth factor receptorinhibitors, in 2010 for subependymal giant cell astrocytoma (SEGA)associated with tuberous sclerosis (TS) in patients who require therapybut are not candidates for surgical resection, and in 2011 forprogressive neuroendocrine tumors of pancreatic origin (PNET) inpatients with unresectable, locally advanced or metastatic disease. Theinteresting but limited clinical success of these mTORC1 compoundsdemonstrates the usefulness of mTOR inhibitors in the treatment ofcancer and transplant rejection, and the increased potential forcompounds with both mTORC1 and mTORC2 inhibitory activity.

The preparation and selection of a solid form of a pharmaceuticalcompound are complex, given that a change in the solid form may affect avariety of physical and chemical properties of the compound, which mayin turn provide benefits or drawbacks in processing, formulation,stability, and bioavailability of the compound. Potential pharmaceuticalsolids include crystalline solids and amorphous solids. An amorphoussolid is characterized by a lack of long-range structural order, whereasa crystalline solid is characterized by structural periodicity. Thedesired class of pharmaceutical solids depends upon the specificapplication; an amorphous solid is sometimes selected on the basis of,e.g., an enhanced dissolution profile, while a crystalline solid may bedesirable for properties, such as physical or chemical stability. SeeVippagunta et al., Adv. Drug. Deliv. Rev., 48:3-26 (2001); Yu, Adv.Drug. Deliv. Rev., 48:27-42 (2001).

Whether crystalline or amorphous, potential solid forms of apharmaceutical compound may include single-component solids. Asingle-component solid contains essentially the pharmaceutical compoundin the absence of other compounds. Variety among single-componentcrystalline materials may potentially arise, e.g., from the phenomenonof polymorphism, wherein multiple three-dimensional arrangements existfor a single pharmaceutical compound. See Byrn et al., Solid StateChemistry of Drugs, SSCI, West Lafayette (1999). The importance ofpolymorphs in pharmaceuticals was underscored by the case of Ritonavir,an HIV protease inhibitor that was formulated as soft gelatin capsules.About two years after the product was launched, the unanticipatedprecipitation of a new, less soluble polymorph in the formulationnecessitated the withdrawal of the product from the market until a moreconsistent formulation could be developed. See Chemburkar et al., Org.Process Res. Dev., 4:413-417 (2000).

Notably, it is not possible to predict a priori if crystalline forms ofa compound even exist, let alone how to successfully prepare them (see,e.g., Braga and Grepioni, 2005, “Making crystals from crystals: a greenroute to crystal engineering and polymorphism,” Chem. Commun.:3635-3645(with respect to crystal engineering, if instructions are not veryprecise and/or if other external factors affect the process, the resultcan be unpredictable); Jones et al., 2006, Pharmaceutical Cocrystals: AnEmerging Approach to Physical Property Enhancement,” MRS Bulletin31:875-879 (At present it is not generally possible to computationallypredict the number of observable polymorphs of even the simplestmolecules); Price, 2004, “The computational prediction of pharmaceuticalcrystal structures and polymorphism,” Advanced Drug Delivery Reviews56:301-319 (“Price”); and Bernstein, 2004, “Crystal Structure Predictionand Polymorphism,” ACA Transactions 39:14-23 (a great deal still needsto be learned and done before one can state with any degree ofconfidence the ability to predict a crystal structure, much lesspolymorphic forms)). The preparation of solid forms is of greatimportance in the development of a safe, effective, stable, andmarketable pharmaceutical compound.

Citation or identification of any references in this disclosure is notto be construed as an admission that the references are prior art tothis disclosure.

3. SUMMARY

Provided herein are compositions of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof. In one embodiment, the solid form is crystalline. Inanother embodiment, the solid form is a single-component crystallineform of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.In yet another embodiment, the solid form is crystalline Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In yet another embodiment, the solid form is a hydrate. In yet anotherembodiment, the solid form is hydrate Form B of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In yet another embodiment, the solid form is anhydrous. In yet anotherembodiment, the solid form is anhydrous Form C of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In yet another embodiment, the solid form is a solvate. In yet anotherembodiment, the solid form is methanol solvate Form D of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In yet another embodiment, the solid form is p-xylene solvate Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In yet another embodiment, the solid form is a pinacol co-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In another embodiment, the isotopologue is enriched in ¹³C, ¹⁴C, ²H, ³Hand/or ¹⁵N. In another embodiment, the isotopologue is enriched in ¹³C,¹⁴C, and/or ²H.

Without intending to be limited by any particular theory, a solid formprovided herein has particular advantageous physical and/or chemicalproperties making them useful, e.g., for manufacturing, processing,formulation and/or storage, while also possessing particularlyadvantageous biological properties, such as, e.g., bioavailabilityand/or biological activity.

Also provided herein are pharmaceutical compositions comprising7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, and one or more pharmaceutically acceptable excipients.

In one embodiment, the pharmaceutical composition comprises a solid formof7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,and one or more pharmaceutically acceptable excipients.

In one embodiment, the pharmaceutical composition comprises Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,and one or more pharmaceutically acceptable excipients

In one embodiment, the pharmaceutical composition comprises Form B of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,and one or more pharmaceutically acceptable excipients.

In one embodiment, the pharmaceutical composition comprises Form C of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,and one or more pharmaceutically acceptable excipients.

In one embodiment, the pharmaceutical composition comprises Form D of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,and one or more pharmaceutically acceptable excipients.

In one embodiment, the pharmaceutical composition comprises Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,and one or more pharmaceutically acceptable excipients.

In one embodiment, the pharmaceutical composition comprises a pinacolco-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,and one or more pharmaceutically acceptable excipients.

In one embodiment, the pharmaceutical composition comprises amorphous7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,and one or more pharmaceutically acceptable excipients.

In another embodiment, the pharmaceutical composition comprises anisotopologue of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,and one ore more pharmaceutically acceptable excipients. In oneembodiment, the isotopologue is enriched in ¹³C, ¹⁴C, ²H, ³H and/or ¹⁵N.In another embodiment, the isotopologue is enriched in ¹³C, ¹⁴C, and/or²H.

Additionally, provided herein are isotopologues of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-oneitself, including isotopologues enriched in ¹³C, ¹⁴C, ²H, ³H and/or ¹⁵N,including those set forth herein.

Additionally, provided herein is are methods of treating or preventing adisease, disorder, or condition in a subject, which comprisesadministering to the subject a therapeutically effective amount of acomposition of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof. In certain embodiments, the disease, disorder, orcondition is cancer, inflammatory conditions, immunological conditions,neurodegenerative diseases, diabetes, obesity, neurological disorders,age-related diseases, and/or cardiovascular conditions, and/orconditions treatable or preventable by inhibition of a kinase pathway.In one embodiment, the kinase pathway is the mTOR/PI3K/Akt pathway.

Provided herein are methods of treating or preventing a disease,disorder, or condition in a subject, which comprise inhibiting a kinasepathway in said subject with a metabolite of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.In certain embodiments, the metabolite is the O-desmethyl metabolite(having the name1-((trans)-4-hydroxycyclohexyl)-7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one).In certain embodiments, the disease, disorder, or condition is cancer,inflammatory conditions, immunological conditions, neurodegenerativediseases, diabetes, obesity, neurological disorders, age-relateddiseases, and/or cardiovascular conditions, and/or conditions treatableor preventable by inhibition of a kinase pathway. In one embodiment, thekinase pathway is the mTOR/PI3K/Akt pathway.

Provided herein are methods of treating or preventing a disease,disorder, or condition in a subject, which comprise administering aneffective amount of a compound that provides a metabolite of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-oneupon administration to said patient. In certain embodiments, themetabolite is the O-desmethyl metabolite (having the name1-((trans)-4-hydroxycyclohexyl)-7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one).In certain embodiments, the disease, disorder, or condition is cancer,inflammatory conditions, immunological conditions, neurodegenerativediseases, diabetes, obesity, neurological disorders, age-relateddiseases, and/or cardiovascular conditions, and/or conditions treatableor preventable by inhibition of a kinase pathway. In one embodiment, thekinase pathway is the mTOR/PI3K/Akt pathway.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form B of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form C of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form D of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of a pinacol co-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of amorphous7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In another embodiment, the method comprises administering to the subjecta therapeutically effective amount of an isotopologue of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.In one embodiment, the isotopologue is enriched in ¹³C, ¹⁴C, ²H, ³Hand/or ¹⁵N. In another embodiment, the isotopologue is enriched in ¹³C,¹⁴C, and/or ²H.

Further provided herein is are methods of treating or preventing aproliferative disease in a subject, which comprises administering to thesubject a therapeutically effective amount of a composition of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form B of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form C of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form D of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of a pinacol co-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount amorphous7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In another embodiment, the method comprises administering to the subjecta therapeutically effective amount of an isotopologue of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.In one embodiment, the isotopologue is enriched in ¹³C, ¹⁴C, ²H, ³Hand/or ¹⁵N. In another embodiment, the isotopologue is enriched in ¹³C,¹⁴C, and/or ²H.

Provided herein are methods of treating or preventing an mTOR-mediateddisease in a subject, which comprises administering to the subject atherapeutically effective amount of a composition of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form B of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form C of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form D of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of a pinacol co-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of amorphous7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In another embodiment, the method comprises administering to the subjecta therapeutically effective amount of an isotopologue of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.In one embodiment, the isotopologue is enriched in ¹³C, ¹⁴C, ²H, ³Hand/or ¹⁵N. In another embodiment, the isotopologue is enriched in ¹³C,¹⁴C, and/or ²H.

Provided herein are methods of inhibiting the growth of a cell,comprising contacting the cell with a composition of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof.

In one embodiment, the method comprises contacting the cell with Form Aof7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises contacting the cell with Form Bof7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises contacting the cell with Form Cof7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises contacting the cell with Form Dof7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises contacting the cell with Form Eof7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises contacting the cell with apinacol co-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises contacting the cell withamorphous7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In another embodiment, the method comprises contacting a cell with anisotopologue of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.In one embodiment, the isotopologue is enriched in ¹³C, ¹⁴C, ²H, ³Hand/or ¹⁵N. In another embodiment, the isotopologue is enriched in ¹³C,¹⁴C, and/or ²H.

Provided herein are methods of modulating the activity of TOR kinase,comprising contacting TOR kinase with a composition of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite, orsolid form thereof.

In one embodiment, the method comprises contacting TOR kinase with FormA of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises contacting TOR kinase with FormB of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises contacting TOR kinase with FormC of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises contacting TOR kinase with FormD of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises contacting TOR kinase with FormE of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises contacting TOR kinase with apinacol co-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises contacting TOR kinase withamorphous7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one.

In another embodiment, the method comprises the method comprisescontacting TOR kinase with an isotopologue of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.In one embodiment, the isotopologue is enriched in ¹³C, ¹⁴C, ²H, ³Hand/or ¹⁵N. In another embodiment, the isotopologue is enriched in ¹³C,¹⁴C, and/or ²H.

Provided herein are methods for treating or preventing a solid tumor,non-Hodgkin lymphoma or multiple myeloma comprising administering aneffective amount of a composition of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,isotopologue, metabolite or a pharmaceutically acceptable salt or solidform thereof, to a subject having a solid tumor, non-Hodgkin lymphoma ormultiple myeloma.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form B of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form C of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form D of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of a pinacol co-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of amorphous7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In another embodiment, the method comprises administering to the subjecta therapeutically effective amount of an isotopologue of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.In one embodiment, the isotopologue is enriched in ¹³C, ¹⁴C, ²H, ³Hand/or ¹⁵N. In another embodiment, the isotopologue is enriched in ¹³C,¹⁴C, and/or ²H.

In certain embodiments, provided herein are methods for achieving aResponse Evaluation Criteria in Solid Tumors (RECIST 1.1) of completeresponse, partial response or stable disease, improving theInternational Workshop Criteria (IWC) for NHL, International UniformResponse Criteria for Multiple Myeloma (IURC), Eastern CooperativeOncology Group Performance Status (ECOG) or Response Assessment forNeuro-Oncology (RANO) Working Group for GBM comprising administering aneffective amount of a composition of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof to a subject having a solid tumor, non-Hodgkin lymphoma ormultiple myeloma

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form B of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form C of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form D of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of a pinacol co-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In one embodiment, the method comprises administering to the subject atherapeutically effective amount of amorphous7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In another embodiment, the method comprises administering to the subjecta therapeutically effective amount of an isotopologue of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.In one embodiment, the isotopologue is enriched in ¹³C, ¹⁴C, ²H, ³Hand/or ¹⁵N. In another embodiment, the isotopologue is enriched in ¹³C,¹⁴C, and/or ²H.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving amorphous7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-onein toluene, MTBE (methyl tert-butyl ether), DIPE (diisopropyl ether),THF (tetrahydrofuran), DME (dimethoxyethane), IPAc (isopropyl acetate),EtOAc (ethyl acetate), MIBK (methyl isobutyl ketone), acetone, IPA(isopropyl alcohol), ethanol, ACN (acetonitrile), nitromethane, orIPA:water (95:5) and allowing the resulting solution to evaporate atroom temperature.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT (butylated hydroxytoluene), IPA and water, heatingand then cooling to room temperature.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT and MeOAc (methyl acetate), heating, cooling to roomtemperature, distilling under vacuum and contacting with n-heptane.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT and MeOAc (methyl acetate), heating, filtering,cooling, distilling under vacuum and contacting with n-heptane.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-oneand7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onepinacol cocrystal in IPA.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT, THF and water, contacting with IPAc, heating,distilling, cooling, contacting with IPAc, heating, distilling and thencooling to room temperature.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT, THF and water, contacting with IPAc, heating,distilling under reduced pressure, contacting with IPAc, and thencooling to room temperature. In one embodiment, the methods additionallycomprise treating with activated carbon.

In certain embodiments, provided herein are methods for making Form B of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT, IPA and water, heating mixture and adding water,cooling the mixture, collecting by filtration, washing with IPA & water,and drying. In certain embodiments, this method further comprises addinga small amount of Form B in water to the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein BHT, IPA and water.

In certain embodiments, provided herein are methods for making Form C of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT, MeOH, distilling to remove MeOH, furtherdistillation with IPA, cooling the mixture, collecting by filtration,washing with IPA and drying.

In certain embodiments, provided herein are methods for making Form D of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT in MeOH, heating, then cooling with stirring,collection by filtration, washing and drying.

In certain embodiments, provided herein are methods for making Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising obtaining a slurry of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-oneinp-xylene/acetone (50/50), heating the slurry to about 60° C.,filtering the slurry to yield a solution, cooling down the solution toabout 25° C. to yield solids, and collecting the solids.

In certain embodiments, provided herein are methods for making a pinacolco-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,comprising mixing7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onewith pinacol in solution, heating until solids are dissolved, distillingsaid solution and seeding with a pinacol co-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

In certain embodiments, provided herein are methods for preparing acomposition provided herein, comprising: (i) weighing out the desiredamount of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the desired amount of excipients; (ii) mixing orblending7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the excipients; (iii) passing the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and excipients through a screen; (iv) mixing or blending7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the excipients; (v) weighing out the desired amount oflubricating agents; (vi) passing the lubricating agents through ascreen; (vii) mixing or blending7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents; (viii)compressing the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents; and (ix)coating the compressed mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents.

In certain embodiments, provided herein are methods for preparing acomposition provided herein, comprising: (i) weighing out the desiredamount of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the desired amount of excipients; (ii) passing theexcipients through a screen; (iii) mixing or blending7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the excipients; (iv) passing the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and excipients through a screen; (v) mixing or blending7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the excipients; (vi) weighing out the desired amount oflubricating agents; (vii) passing the lubricating agents through ascreen; (viii) mixing or blending7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents; (ix)compressing the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents; and (x) coatingthe compressed mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents.

4. BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts an X-ray powder diffractogram of Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 2 depicts polar light microscopy photographs of Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 3 depicts a thermogravimetric thermogram (top) and a differentialscanning calorimetric thermogram (bottom) of Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 4 depicts kinetic (top) and isotherm (bottom) DVS curves of Form Aof7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 5 depicts dissolution profiles of 20 mg tablets of Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one(Core vs Coated).

FIG. 6 depicts a differential scanning calorimetric (DSC) thermogram ofa pinacol co-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 7 depicts an X-ray powder diffractogram of a pinacol co-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 8 depicts plasma concentration-time profiles in healthy adult malesadministered a single 20 mg oral dose of Compound A.

FIG. 9 depicts an X-ray powder diffractogram of Form B of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 10 depicts a differential scanning calorimetric (DSC) thermogram ofForm B of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 11 depicts an X-ray powder diffractogram of Form C of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 12 depicts a differential scanning calorimetric (DSC) thermogram ofForm C of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 13 depicts a differential scanning calorimetric (DSC) thermogram ofForm D of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 14 depicts an X-ray powder diffractogram of Form D of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 15 depicts an X-ray powder diffractogram stack plot of Form A, FormE as a wet solid, Form E as a dried solid, Form E as a wet solid afterexposure to accelerated aging conditions (AAC) and Form E as a driedsolid after exposure to AAC of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one(from bottom to top).

FIG. 16 depicts a crystal packing pattern of Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 17: (A) depicts a digital image of Form E as a wet solid of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.(B) depicts a digital image of Form E as a dried solid of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.(C) depicts a digital image of Form E as a wet solid after exposure toAAC of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.(D) depicts a digital image of Form E as a dried solid after exposure toAAC of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 18 depicts a thermogravimetrical analysis and single differentialthermal analysis of Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 19 depicts a thermogravimetric analysis coupled with massspectroscopy of Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

FIG. 20 depicts high performance liquid chromatography coupled with massspectrometry of Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.

5. DETAILED DESCRIPTION 5.1 Definitions

To facilitate understanding of the disclosure set forth herein, a numberof terms are defined below.

Generally, the nomenclature used herein and the laboratory procedures inorganic chemistry, medicinal chemistry, and pharmacology describedherein are those well known and commonly employed in the art. Unlessdefined otherwise, all technical and scientific terms used hereingenerally have the same meaning as commonly understood by one ofordinary skill in the art to which this disclosure belongs.

It should be noted that if there is a discrepancy between a depictedstructure and a name given that structure, the depicted structure is tobe accorded more weight. In addition, if the stereochemistry of astructure or a portion of a structure is not indicated with, forexample, bold or dashed lines, the structure or portion of the structureis to be interpreted as encompassing all stereoisomers of it.

The term “Compound A” refers to7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,also having the chemical names7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((1r,4r)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-oneand7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((1R*,4R*)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,which has the following structure:

including pharmaceutically acceptable salts, isotopologues, solid formsand metabolites thereof.

Compound A can be prepared according to the methods described in U.S.Pat. Appl. Publ. Nos. 2010/0216781 and 2011/0137028, the disclosure ofeach of which is incorporated herein by reference in its entirety.Compound A can also be synthesized according to other methods apparentto those of skill in the art based upon the teaching herein.

The term “subject” refers to an animal, including, but not limited to, aprimate (e.g., human), cow, pig, sheep, goat, horse, dog, cat, rabbit,rat, or mouse. The terms “subject” and “patient” are used hereininterchangeably in reference, for example, to a mammalian subject, suchas a human subject, in one embodiment, a human. In one embodiment, thesubject has or is susceptible to having a disease, disorder, orcondition provided herein.

The term “treat,” “treating,” or “treatment” means an alleviation, inwhole or in part, of a disease, disorder, or condition provided herein,or one or more symptoms associated with the disease, disorder, orcondition, or slowing, or halting of further progression or worsening ofthe disease, disorder, or condition, or one or more symptoms associatedwith the disease, disorder, or condition.

The term “prevent,” “preventing,” or “prevention” means prevention ofthe onset, recurrence, or spread of a disease, disorder, or conditionprovided herein, or one or more symptoms associated with the disease,disorder, or condition, in a subject at risk for developing the disease,disorder, or condition.

The term “effective amount” or “therapeutically effective amount” refersto, in one embodiment, an amount of Compound A capable of alleviating,in whole or in part, one or more symptoms associated with a disease,disorder, or condition provided herein, or slowing or halting furtherprogression or worsening of one or more of the symptoms of the disease,disorder, or condition; in another embodiment, an amount capable ofpreventing or providing prophylaxis for the disease, disorder, orcondition in a subject at risk for developing the disease, disorder, orcondition, such as cancer, inflammatory conditions, immunologicalconditions, neurodegenerative diseases, diabetes, obesity, neurologicaldisorders, age-related diseases, and/or cardiovascular conditions,and/or diseases, disorders, and conditions treatable or preventable byinhibition of a kinase pathway, for example, the mTOR/PI3K/Akt pathway.In one embodiment, an effective amount of a compound is an amount thatinhibits a kinase in a cell, such as, for example, in vitro or in vivo.In one embodiment the kinase is TOR kinase. In certain embodiments, theeffective amount of a compound inhibits the kinase in a cell by about10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%,about 80%, about 90%, or about 99%, compared to the activity of thekinase in an untreated cell. In one embodiment, “effective amount”refers to the amount of Compound A capable of alleviating, in whole orin part, symptoms associated with a solid tumor (for example, aneuroendocrine tumor, non-small cell lung cancer, glioblastomamultiforme, hepatocellular carcinoma, breast cancer, colorectal cancer,salivary cancer, pancreatic cancer, adenocystic cancer, adrenal cancer,esophageal cancer, renal cancer, leiomyosarcoma, or paraganglioma,including advanced solid tumors), non-Hodgkin lymphoma or multiplemyeloma, or slowing or halting further progression or worsening of thosesymptoms, or treating or preventing a solid tumor (for example, aneuroendocrine tumor, non-small cell lung cancer, glioblastomamultiforme, hepatocellular carcinoma, breast cancer, colorectal cancer,salivary cancer, pancreatic cancer, adenocystic cancer, adrenal cancer,esophageal cancer, renal cancer, leiomyosarcoma, or paraganglioma),non-Hodgkin lymphoma or multiple myeloma in a subject having or at riskfor having a solid tumor, non-Hodgkin lymphoma or multiple myeloma. Aswill be apparent to those skilled in the art, it is to be expected thatthe effective amount of a compound disclosed herein may vary dependingon the indication being treated, e.g., the effective amount of thecompound would likely be different for treating patients suffering from,or at risk for, inflammatory conditions relative to the effective amountof the compound for treating patients suffering from, or at risk of, adifferent disorder, e.g., a disorder provided herein.

In the context of a solid tumor (for example, a neuroendocrine tumor,non-small cell lung cancer, glioblastoma multiforme, hepatocellularcarcinoma, breast cancer, colorectal cancer, salivary cancer, pancreaticcancer, adenocystic cancer, adrenal cancer, esophageal cancer, renalcancer, leiomyosarcoma, or paraganglioma, including advanced solidtumors), non-Hodgkin lymphoma or multiple myeloma, inhibition may beassessed by inhibition or retarding of disease progression, inhibitionof tumor growth, reduction or regression of primary and/or secondarytumor (s), relief of tumor-related symptoms, improvement in quality oflife, inhibition of tumor secreted factors (including tumor secretedhormones, such as those that contribute to carcinoid syndrome),reductions in endocrine hormone markers (for example, chromogranin,gastrin, serotonin, and/or glucagon), delayed appearance or recurrenceof primary or secondary tumors, slowed development of primary and/orsecondary tumors, decreased occurrence of primary and/or secondarytumors, slowed or decreased severity of secondary effects of disease,arrested tumor growth and/or regression of tumors, increased Time ToProgression (TTP), increased Progression Free Survival (PFS), increasedOverall Survival (OS), among others. OS as used herein means the timefrom randomization until death from any cause, and is measured in theintent-to-treat population. TTP as used herein means the time fromrandomization until objective tumor progression; TTP does not includedeaths. As used herein, PFS means the time from randomization untilobjective tumor progression or death. In one embodiment, PFS rates willbe computed using the Kaplan-Meier estimates. In the extreme, completeinhibition, is referred to herein as prevention or chemoprevention. Inthis context, the term “prevention” includes either preventing the onsetof clinically evident solid tumor (for example, a neuroendocrine tumor,non-small cell lung cancer, glioblastoma multiforme, hepatocellularcarcinoma, breast cancer, colorectal cancer, salivary cancer, pancreaticcancer, adenocystic cancer, adrenal cancer, esophageal cancer, renalcancer, leiomyosarcoma, or paraganglioma, including advanced solidtumors), non-Hodgkin lymphoma or multiple myeloma altogether orpreventing the onset of a preclinically evident stage of a solid tumor(for example, a neuroendocrine tumor, non-small cell lung cancer,glioblastoma multiforme, hepatocellular carcinoma, breast cancer,colorectal cancer, salivary cancer, pancreatic cancer, adenocysticcancer, adrenal cancer, esophageal cancer, renal cancer, leiomyosarcoma,or paraganglioma, including advanced solid tumors), non-Hodgkin lymphomaor multiple myeloma. Also intended to be encompassed by this definitionis the prevention of transformation into malignant cells or to arrest orreverse the progression of premalignant cells to malignant cells. Thisincludes prophylactic treatment of those at risk of developing a solidtumor (for example, a neuroendocrine tumor, non-small cell lung cancer,glioblastoma multiforme, hepatocellular carcinoma, breast cancer,colorectal cancer, salivary cancer, pancreatic cancer, adenocysticcancer, adrenal cancer, esophageal cancer, renal cancer, leiomyosarcoma,or paraganglioma, including advanced solid tumors), non-Hodgkin lymphomaor multiple myeloma.

The term “cancer” refers to any of various malignant neoplasmscharacterized by the proliferation of cells that can invade surroundingtissue and metastasize to new body sites. Both benign and malignanttumors are classified according to the type of tissue in which they arefound. For example, fibromas are neoplasms of fibrous connective tissue,and melanomas are abnormal growths of pigment (melanin) cells. Malignanttumors originating from epithelial tissue, e.g., in skin, bronchi, andstomach, are termed carcinomas. Malignancies of epithelial glandulartissue such as are found in the breast, prostate, and colon, are knownas adenocarcinomas. Malignant growths of connective tissue, e.g.,muscle, cartilage, lymph tissue, and bone, are called sarcomas.Lymphomas and leukemias are malignancies arising among white bloodcells. Through the process of metastasis, tumor cell migration to otherareas of the body establishes neoplasms in areas away from the site ofinitial appearance. Bone tissues are one of the most favored sites ofmetastases of malignant tumors, occurring in about 30% of all cancercases. Among malignant tumors, cancers of the lung, breast, prostate orthe like are particularly known to be likely to metastasize to bone.

An “advanced solid tumor” as used herein, means a solid tumor that hasspread locally, or metastasized or spread to another part of the body.

In certain embodiments, the treatment may be assessed by ResponseEvaluation Criteria in Solid Tumors (RECIST 1.1) (see Thereasse P., etal. New Guidelines to Evaluate the Response to Treatment in SolidTumors. J. of the National Cancer Institute; 2000; (92) 205-216 andEisenhauer E. A., Thereasse P., Bogaerts J., et al. New responseevaluation criteria in solid tumours: Revised RECIST guideline (version1.1). European J. Cancer; 2009; (45) 228-247). Overall responses for allpossible combinations of tumor responses in target and non-targetlesions with our without the appearance of new lesions are as follows:

Target lesions Non-target lesions New lesions Overall response CR CR NoCR CR Incomplete No PR response/SD PR Non-PD No PR SD Non-PD No SD PDAny Yes or no PD Any PD Yes or no PD Any Any Yes PD CR = completeresponse; PR = partial response; SD = stable disease; and PD =progressive disease.

With respect to the evaluation of target lesions, complete response (CR)is the disappearance of all target lesions, partial response (PR) is atleast a 30% decrease in the sum of the longest diameter of targetlesions, taking as reference the baseline sum longest diameter,progressive disease (PD) is at least a 20% increase in the sum of thelongest diameter of target lesions, taking as reference the smallest sumlongest diameter recorded since the treatment started or the appearanceof one or more new lesions and stable disease (SD) is neither sufficientshrinkage to qualify for partial response nor sufficient increase toqualify for progressive disease, taking as reference the smallest sumlongest diameter since the treatment started.

With respect to the evaluation of non-target lesions, complete response(CR) is the disappearance of all non-target lesions and normalization oftumor marker level; incomplete response/stable disease (SD) is thepersistence of one or more non-target lesion(s) and/or the maintenanceof tumor marker level above the normal limits, and progressive disease(PD) is the appearance of one or more new lesions and/or unequivocalprogression of existing non-target lesions.

In certain embodiments, the treatment of lymphoma may be assessed by theInternational Workshop Criteria (IWC) for non-Hodgkin lymphoma (NHL)(see Cheson B D, Pfistner B, Juweid, M E, et. al. Revised ResponseCriteria for Malignant Lymphoma. J. Clin. Oncol: 2007: (25) 579-586),using the response and endpoint definitions shown below:

Response Definition Nodal Masses Spleen, liver Bone Marrow CRDisappearance (a) FDG-avid or PET Not palpable, Infiltrate of allevidence positive prior to therapy; nodules cleared on of disease massof any size permitted disappeared repeat biopsy; if PET negative ifindeterminate (b) Variably FDG-avid or by morphology, PET negative;regression immunohisto- to normal size on CT chemistry should benegative PR Regression of ≥50% decrease in SPD of ≥50% Irrelevant ifmeasurable up to 6 largest dominant decrease in positive prior diseaseand no masses; no increase in size SPD of to therapy; cell new sites ofother nodes nodules (for type should be (a) FDG-avid or PET singlenodule specified positive prior to therapy; in greatest one or more PETpositive transverse at previously involved site diameter); no (b)Variably FDG-avid or increase in PET negative; regression size of liveron CT or spleen SD Failure to (a) FDG-avid or PET attain CR/PR positiveprior to therapy; or PD PET positive at prior sites of disease and nonew sites on CT or PET (b) Variably FDG-avid or PET negative; no changein size of previous lesions on CT PD or Any new Appearance of a new ≥50%New or relapsed lesion or lesion(s) ≥1.5 cm in any increase recurrentdisease increase by ≥50% axis, ≥50% increase in from nadir ininvolvement of previously SPD of more than one node, the SPD of involvedsites or ≥50% increase in any previous from nadir longest diameter of alesions previously identifed node ≥1 cm in short axis Lesions PETpositive if FDG-avid lymphoma or PET positive prior to therapyAbbreviations: CR, complete remission; FDG, [¹⁸F]fluorodeoxyglucose;PET, positron emission tomography; CT, computed tomography; PR, partialremission; SPD, sum of the product of the diameters; SD, stable disease;PD, progressive disease.

Pa- End point tients Definition Measured from Primary Overall survivalAll Death as a result of any Entry onto cause study Progression-free AllDisease progression or Entry onto survival death as a result of anystudy cause Secondary Event-free All Failure of treatment or Entry ontosurvival death as result of any study cause Time to All Time toprogression or Entry onto progression death as a result of studylymphoma Disease-free In CR Time to relapse or death Documentationsurvival as a result of lymphoma of response or acute toxicity oftreatment Response In CR Time to relapse or Documentation duration or PRprogression of response Lymphoma- All Time to death as a result Entryonto specific of lymphoma study survival Time to next All Time to newtreatment End of primary treatment treatment Abbreviations: CR: completeremission; PR: partial remission.

In one embodiment, the end point for lymphoma is evidence of clinicalbenefit. Clinical benefit may reflect improvement in quality of life, orreduction in patient symptoms, transfusion requirements, frequentinfections, or other parameters. Time to reappearance or progression oflymphoma-related symptoms can also be used in this end point.

In certain embodiments, the treatment of multiple myeloma may beassessed by the International Uniform Response Criteria for MultipleMyeloma (IURC) (see Durie BGM, Harousseau J-L, Miguel J S, et al.International uniform response criteria for multiple myeloma. Leukemia,2006; (10) 10: 1-7), using the response and endpoint definitions shownbelow:

Response Subcategory Response Criteria^(a) sCR CR as defined below plusNormal FLC ratio and Absence of clonal cells in bone marrow^(b) byimmunohistochemistry or immunofluorescence^(c) CR Negativeimmunofixation on the serum and urine and Disappearance of any softtissue plasmacytomas and <5% plasma cells in bone marrow^(b) VGPR Serumand urine M-protein detectable by immunofixation but not onelectrophoresis or 90% or greater reduction in serum M-protein plusurine M-protein level <100 mg per 24 h PR ≥50% reduction of serumM-protein and reduction in 24-h urinary M-protein by ≥90% or to <200 mgper 24 h If the serum and urine M-protein are unmeasurable,^(d) a ≥50%decrease in the difference between involved and uninvolved FLC levels isrequired in place of the M-protein criteria If serum and urine M-proteinare unmeasurable, and serum free light assay is also unmeasurable, ≥50%reduction in plasma cells is required in place of M-protein, providedbaseline bone marrow plasma cell percentage was ≥30% In addition to theabove listed criteria, if present at baseline, a ≥50% reduction in thesize of soft tissue plasmacytomas is also required SD (not recommendedfor use Not meeting criteria for CR, VGPR, PR or progressive disease asan indicator of response; stability of disease is best described byproviding the time to progression estimates) Abbreviations: CR, completeresponse; FLC, free light chain; PR, partial response; SD, stabledisease; sCR, stringent complete response; VGPR, very good partialresponse; ^(a)All response categories require two consecutiveassessments made at anytime before the institution of any new therapy;all categories also require no known evidence of progressive or new bonelesions if radiographic studies were performed. Radiographic studies arenot required to satisfy these response requirements; ^(b)Confirmationwith repeat bone marrow biopsy not needed; ^(c)Presence/absence ofclonal cells is based upon the κ/λ ratio. An abnormal κ/λ ratio byimmunohistochemistry and/or immunofluorescence requires a minimum of 100plasma cells for analysis. An abnormal ratio reflecting presence of anabnormal clone is κ/λ of >4:1 or <1:2. ^(d)Measurable disease defined byat least one of the following measurements: Bone marrow plasma cells≥30%; Serum M-protein ≥1 g/dl (≥10 gm/l)[10 g/l]; Urine M-protein ≥200mg/24 h; Serum FLC assay: Involved FLC level ≥10 mg/dl (≥100 mg/l);provided serum FLC ratio is abnormal.

The procedures, conventions, and definitions described below provideguidance for implementing the recommendations from the ResponseAssessment for Neuro-Oncology (RANO) Working Group regarding responsecriteria for high-grade gliomas (Wen P., Macdonald, D R., Reardon, D A.,et al. Updated response assessment criteria for highgrade gliomas:Response assessment in neuro-oncology working group. J Clin Oncol 2010;28: 1963-1972). Primary modifications to the RANO criteria for Criteriafor Time Point Responses (TPR) can include the addition of operationalconventions for defining changes in glucocorticoid dose, and the removalof subjects' clinical deterioration component to focus on objectiveradiologic assessments. The baseline MRI scan is defined as theassessment performed at the end of the post-surgery rest period, priorto re-initiating compound treatment. The baseline MRI is used as thereference for assessing complete response (CR) and partial response(PR). Whereas, the smallest SPD (sum of the products of perpendiculardiameters) obtained either at baseline or at subsequent assessments willbe designated the nadir assessment and utilized as the reference fordetermining progression. For the 5 days preceding any protocol-definedMRI scan, subjects receive either no glucocorticoids or are on a stabledose of glucocorticoids. A stable dose is defined as the same daily dosefor the 5 consecutive days preceding the MRI scan. If the prescribedglucocorticoid dose is changed in the 5 days before the baseline scan, anew baseline scan is required with glucocorticoid use meeting thecriteria described above. The following definitions will be used.

Measurable Lesions: Measurable lesions are contrast-enhancing lesionsthat can be measured bidimensionally. A measurement is made of themaximal enhancing tumor diameter (also known as the longest diameter,LD). The greatest perpendicular diameter is measured on the same image.The cross hairs of bidimensional measurements should cross and theproduct of these diameters will be calculated.

Minimal Diameter: T1-weighted image in which the sections are 5 mm with1 mm skip. The minimal LD of a measurable lesion is set as 5 mm by 5 mm.Larger diameters may be required for inclusion and/or designation astarget lesions. After baseline, target lesions that become smaller thanthe minimum requirement for measurement or become no longer amenable tobidimensional measurement will be recorded at the default value of 5 mmfor each diameter below 5 mm. Lesions that disappear will be recorded as0 mm by 0 mm.

Multicentric Lesions: Lesions that are considered multicentric (asopposed to continuous) are lesions where there is normal interveningbrain tissue between the two (or more) lesions. For multicentric lesionsthat are discrete foci of enhancement, the approach is to separatelymeasure each enhancing lesion that meets the inclusion criteria. Ifthere is no normal brain tissue between two (or more) lesions, they willbe considered the same lesion.

Nonmeasurable Lesions: All lesions that do not meet the criteria formeasurable disease as defined above will be considered non-measurablelesions, as well as all nonenhancing and other truly nonmeasurablelesions. Nonmeasurable lesions include foci of enhancement that are lessthan the specified smallest diameter (ie., less than 5 mm by 5 mm),nonenhancing lesions (eg., as seen on T1-weighted post-contrast,T2-weighted, or fluid-attenuated inversion recovery (FLAIR) images),hemorrhagic or predominantly cystic or necrotic lesions, andleptomeningeal tumor. Hemorrhagic lesions often have intrinsicT1-weighted hyperintensity that could be misinterpreted as enhancingtumor, and for this reason, the pre-contrast T1-weighted image may beexamined to exclude baseline or interval sub-acute hemorrhage.

At baseline, lesions will be classified as follows: Target lesions: Upto 5 measurable lesions can be selected as target lesions with eachmeasuring at least 10 mm by 5 mm, representative of the subject'sdisease; Non-target lesions: All other lesions, including allnonmeasurable lesions (including mass effects and T2/FLAIR findings) andany measurable lesion not selected as a target lesion. At baseline,target lesions are to be measured as described in the definition formeasurable lesions and the SPD of all target lesions is to bedetermined. The presence of all other lesions is to be documented. Atall post-treatment evaluations, the baseline classification of lesionsas target and non-target lesions will be maintained and lesions will bedocumented and described in a consistent fashion over time (eg.,recorded in the same order on source documents and eCRFs). Allmeasurable and nonmeasurable lesions must be assessed using the sametechnique as at baseline (e.g., subjects should be imaged on the sameMRI scanner or at least with the same magnet strength) for the durationof the study to reduce difficulties in interpreting changes. At eachevaluation, target lesions will be measured and the SPD calculated.Non-target lesions will be assessed qualitatively and new lesions, ifany, will be documented separately. At each evaluation, a time pointresponse will be determined for target lesions, non-target lesions, andnew lesion. Tumor progression can be established even if only a subsetof lesions is assessed. However, unless progression is observed,objective status (stable disease, PR or CR) can only be determined whenall lesions are assessed.

Confirmation assessments for overall time point responses of CR and PRwill be performed at the next scheduled assessment, but confirmation maynot occur if scans have an interval of <28 days. Best response,incorporating confirmation requirements, will be derived from the seriesof time points.

The term “contacting” or “contact” is meant to refer to bringingtogether of a therapeutic agent and cell or tissue such that aphysiological and/or chemical effect takes place as a result of suchcontact. Contacting can take place in vitro, ex vivo, or in vivo. In oneembodiment, a therapeutic agent is contacted with a cell in cell culture(in vitro) to determine the effect of the therapeutic agent on the cell.In another embodiment, the contacting of a therapeutic agent with a cellor tissue includes the administration of a therapeutic agent to asubject having the cell or tissue to be contacted.

The term “solid form” refers to a physical form which is notpredominantly in a liquid or a gaseous state. As used herein and unlessotherwise specified, the term “solid form,” when used herein to refer toCompound A, refers to a physical form comprising Compound A which is notpredominantly in a liquid or a gaseous state. A solid form may be acrystalline form, an amorphous form, or a mixture thereof. In certainembodiments, a solid form may be a liquid crystal. In certainembodiments, the term “solid forms comprising Compound A” includescrystal forms comprising Compound A, amorphous forms comprising CompoundA, and mixtures thereof.

As used herein and unless otherwise specified, the term “crystalline”when used to describe a compound, substance, modification, material,component or product, unless otherwise specified, means that thecompound, substance, modification, material, component or product issubstantially crystalline as determined by X-ray diffraction. See, e.g.,Remington: The Science and Practice of Pharmacy, 21st edition,Lippincott, Williams and Wilkins, Baltimore, Md. (2005); The UnitedStates Pharmacopeia, 23^(rd) ed., 1843-1844 (1995).

The term “crystal form” or “crystalline form” refers to a solid formthat is crystalline. In certain embodiments, crystal forms includesalts. In certain embodiments, a crystal form of a substance may besubstantially free of amorphous forms and/or other crystal forms. Incertain embodiments, a crystal form of a substance may contain less thanabout 1%, less than about 2%, less than about 3%, less than about 4%,less than about 5%, less than about 6%, less than about 7%, less thanabout 8%, less than about 9%, less than about 10%, less than about 15%,less than about 20%, less than about 25%, less than about 30%, less thanabout 35%, less than about 40%, less than about 45%, or less than about50% by weight of one or more amorphous forms and/or other crystal forms.In certain embodiments, a crystal form of a substance may be physicallyand/or chemically pure. In certain embodiments, a crystal form of asubstance may be about 99%, about 98%, about 97%, about 96%, about 95%,about 94%, about 93%, about 92%, about 91%, or about 90% physicallyand/or chemically pure.

The term “amorphous” or “amorphous form” means that the substance,component, or product in question is not substantially crystalline asdetermined by X-ray diffraction. In particular, the term “amorphousform” describes a disordered solid form, i.e., a solid form lacking longrange crystalline order. In certain embodiments, an amorphous form of asubstance may be substantially free of other amorphous forms and/orcrystal forms. In certain embodiments, an amorphous form of a substancemay contain less than about 1%, less than about 2%, less than about 3%,less than about 4%, less than about 5%, less than about 10%, less thanabout 15%, less than about 20%, less than about 25%, less than about30%, less than about 35%, less than about 40%, less than about 45%, orless than about 50% by weight of one or more other amorphous formsand/or crystal forms on a weight basis. In certain embodiments, anamorphous form of a substance may be physically and/or chemically pure.In certain embodiments, an amorphous form of a substance be about 99%,about 98%, about 97%, about 96%, about 95%, about 94%, about 93%, about92%, about 91%, or about 90% physically and/or chemically pure.

The term “solvate” means a crystalline structure comprised of eitherstoichiometric or nonstoichiometric amounts of a solvent incorporatedwithin the crystalline structure.

The term “hydrate” means a crystalline structure comprised of eitherstoichiometric or nonstoichiometric amounts of water incorporated withinthe crystalline structure.

Techniques for characterizing crystal forms and amorphous forms include,but are not limited to, thermal gravimetric analysis (TGA), differentialscanning calorimetry (DSC), X-ray powder diffractometry (XRPD),single-crystal X-ray diffractometry, vibrational spectroscopy, e.g.,infrared (IR) and Raman spectroscopy, solid-state and solution nuclearmagnetic resonance (NMR) spectroscopy, optical microscopy, hot stageoptical microscopy, scanning electron microscopy (SEM), electroncrystallography and quantitative analysis, particle size analysis (PSA),surface area analysis, solubility measurements, dissolutionmeasurements, elemental analysis, and Karl Fischer analysis.Characteristic unit cell parameters may be determined using one or moretechniques such as, but not limited to, X-ray diffraction and neutrondiffraction, including single-crystal diffraction and powderdiffraction. Techniques useful for analyzing powder diffraction datainclude profile refinement, such as Rietveld refinement, which may beused, e.g., to analyze diffraction peaks associated with a single phasein a sample comprising more than one solid phase. Other methods usefulfor analyzing powder diffraction data include unit cell indexing, whichallows one of skill in the art to determine unit cell parameters from asample comprising crystalline powder.

The term “pharmaceutically acceptable salt(s)” means a salt preparedfrom a pharmaceutically acceptable non-toxic acid or base including aninorganic acid and base and an organic acid and base. Suitablepharmaceutically acceptable base addition salts of Compound A include,but are not limited to metallic salts made from aluminum, calcium,lithium, magnesium, potassium, sodium and zinc or organic salts madefrom lysine, N,N′-dibenzylethylenediamine, chloroprocaine, choline,diethanolamine, ethylenediamine, meglumine (N-methylglucamine) andprocaine. Suitable non-toxic acids include, but are not limited to,inorganic and organic acids such as acetic, alginic, anthranilic,benzenesulfonic, benzoic, camphorsulfonic, citric, ethenesulfonic,formic, fumaric, furoic, galacturonic, gluconic, glucuronic, glutamic,glycolic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic,mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic,phenylacetic, phosphoric, propionic, salicylic, stearic, succinic,sulfanilic, sulfuric, tartaric acid, and p-toluenesulfonic acid.Specific non-toxic acids include hydrochloric, hydrobromic, phosphoric,sulfuric, and methanesulfonic acids. Examples of specific salts thusinclude hydrochloride and mesylate salts. Others are well-known in theart, see for example, Remington's Pharmaceutical Sciences, 18th eds.,Mack Publishing, Easton Pa. (1990) or Remington: The Science andPractice of Pharmacy, 19th eds., Mack Publishing, Easton Pa. (1995).

The term “isotopologue” means any form of Compound A, includingmetabolites thereof, in which at least one atom of natural isotopicabundance is replaced with an isotopically enriched form that differsfrom natural abundance. An isotopologue can be based on replacement ofhydrogen for deuterium and/or tritium. Similarly, naturally abundant ¹²Ccan be replaced with ¹³C or ¹⁴C, naturally abundant ¹⁴N can be replacedwith ¹⁵N, and naturally abundant ¹⁶O with ¹⁷O or ¹⁸O, and so on in anycombination. Other isotopologues can be based on isotopic enrichment offluorine, sulfur, phosphorus, boron, and the like. Isotopologues caninclude replacing any number atoms within the compound with isotopicallyenriched forms. The isotopic enrichment can be effected to any degree,including, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 95,and 99, and 100% enrichment, including any value in between andfractions thereof.

The term “metabolite” means any compound produced upon administration ofCompound A to a subject. In one embodiment, the metabolite of Compound Ais the O-desmethyl metabolite (having the name1-((trans)-4-hydroxycyclohexyl)-7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one),alternatively named1-((1r,4r)-4-hydroxycyclohexyl)-7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,having the structure:

The term “about” or “approximately” means an acceptable error for aparticular value as determined by one of ordinary skill in the art,which depends in part on how the value is measured or determined. Incertain embodiments, the term “about” or “approximately” means within 1,2, 3, or 4 standard deviations. In certain embodiments, the term “about”or “approximately” means within 50%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%,4%, 3%, 2%, 1%, 0.5%, or 0.05% of a given value or range.

As used herein and unless otherwise specified, a sample comprising aparticular crystal form or amorphous form that is “substantially pure,”e.g., substantially free of other solid forms and/or of other chemicalcompounds, contains, in particular embodiments, less than about 25%,less than about 20%, less than about 15%, less than about 10%, less thanabout 9%, less than about 8%, less than about 7%, less than about 6%,less than about 5%, less than about 4%, less than about 3%, less thanabout 2%, less than about 1%, less than about 0.75%, less than about0.5%, less than about 0.25%, or less than about 0.1% by weight of one ormore other solid forms and/or of other chemical compounds.

As used herein and unless otherwise specified, a sample or compositionthat is “substantially free” of one or more other solid forms and/orother chemical compounds means that the composition contains, inparticular embodiments, less than about 25%, less than about 20%, lessthan about 15%, less than about 10%, less than about 9%, less than about8%, less than about 7%, less than about 6%, less than about 5%, lessthan about 4%, less than about 3%, less than about 2%, less than about1%, less than about 0.75%, less than about 0.5%, less than about 0.25%,or less than about 0.1% by weight of one or more other solid formsand/or other chemical compounds.

As used herein, the term “pharmaceutically acceptable salt(s)” refers toa salt prepared from a pharmaceutically acceptable non-toxic acid orbase including an inorganic acid and base and an organic acid and base.Suitable pharmaceutically acceptable base addition salts of Compound Ainclude, but are not limited to metallic salts made from aluminum,calcium, lithium, magnesium, potassium, sodium and zinc or organic saltsmade from lysine, N,N′-dibenzylethylenediamine, chloroprocaine, choline,diethanolamine, ethylenediamine, meglumine (N-methylglucamine) andprocaine. Suitable non-toxic acids include, but are not limited to,inorganic and organic acids such as acetic, alginic, anthranilic,benzenesulfonic, benzoic, camphorsulfonic, citric, ethenesulfonic,formic, fumaric, furoic, galacturonic, gluconic, glucuronic, glutamic,glycolic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic,mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic,phenylacetic, phosphoric, propionic, salicylic, stearic, succinic,sulfanilic, sulfuric, tartaric acid, and p-toluenesulfonic acid.Specific non-toxic acids include hydrochloric, hydrobromic, phosphoric,sulfuric, and methanesulfonic acids. Examples of specific salts thusinclude hydrochloride and mesylate salts. Others are well-known in theart, see for example, Remington's Pharmaceutical Sciences, 18th eds.,Mack Publishing, Easton Pa. (1990) or Remington: The Science andPractice of Pharmacy, 19th eds., Mack Publishing, Easton Pa. (1995).

The term “stereoisomer” or “stereomerically pure” means one stereoisomerof a compound that is substantially free of other stereoisomers of thatcompound. For example, a stereomerically pure compound having one chiralcenter will be substantially free of the opposite enantiomer of thecompound. A stereomerically pure compound having two chiral centers willbe substantially free of other diastereomers of the compound. In certainembodiments, a stereomerically pure compound comprises greater thanabout 80% by weight of one stereoisomer of the compound and less thanabout 20% by weight of other stereoisomers of the compound, greater thanabout 90% by weight of one stereoisomer of the compound and less thanabout 10% by weight of the other stereoisomers of the compound, greaterthan about 95% by weight of one stereoisomer of the compound and lessthan about 5% by weight of the other stereoisomers of the compound, orgreater than about 97% by weight of one stereoisomer of the compound andless than about 3% by weight of the other stereoisomers of the compound.

5.2 Solid Forms of Compound A

In one embodiment, provided herein is a solid form of Compound A or apharmaceutically acceptable salt thereof. In certain embodiments, thesolid form is crystalline. In certain embodiments, the solid form is asingle-component solid form. In certain embodiments, the solid form isanhydrous.

While not intending to be bound by any particular theory, certain solidforms are characterized by physical properties, e.g., stability,solubility and dissolution rate, appropriate for pharmaceutical andtherapeutic dosage forms. Moreover, while not wishing to be bound by anyparticular theory, certain solid forms are characterized by physicalproperties (e.g., density, compressibility, hardness, morphology,cleavage, stickiness, solubility, water uptake, electrical properties,thermal behavior, solid-state reactivity, physical stability, andchemical stability) affecting particular processes (e.g., yield,filtration, washing, drying, milling, mixing, tableting, flowability,dissolution, formulation, and lyophilization) which make certain solidforms suitable for the manufacture of a solid dosage form. Suchproperties can be determined using particular analytical chemicaltechniques, including solid-state analytical techniques (e.g., X-raydiffraction, microscopy, spectroscopy and thermal analysis), asdescribed herein and known in the art.

The solid forms provided herein (for example, Form A of Compound A) maybe characterized using a number of methods known to a person skilled inthe art, including, but not limited to, single crystal X-raydiffraction, X-ray powder diffraction (XRPD), microscopy (e.g., scanningelectron microscopy (SEM)), thermal analysis (e.g., differentialscanning calorimetry (DSC), thermal gravimetric analysis (TGA), andhot-stage microscopy), and spectroscopy (e.g., infrared, Raman, andsolid-state nuclear magnetic resonance). The particle size and sizedistribution of the solid form provided herein may be determined byconventional methods, such as laser light scattering technique.

The purity of the solid forms provided herein may be determined bystandard analytical methods, such as thin layer chromatography (TLC),gel electrophoresis, gas chromatography, high performance liquidchromatography (HPLC), and mass spectrometry (MS).

It should be understood that the numerical values of the peaks of anX-ray powder diffraction pattern may vary slightly from one machine toanother or from one sample to another, and so the values quoted are notto be construed as absolute, but with an allowable variability, such as+0.2 degrees two-theta (degrees two-theta also noted as degrees 2θ, or20) (see United States Pharmacopoeia, page 2228 (2003)).

In one embodiment, provided herein is Form A of Compound A. In oneembodiment, Form A of Compound A has an X-ray powder diffraction patternsubstantially as shown in FIG. 1. In one embodiment, Form A of CompoundA has an X-ray powder diffraction pattern comprised of one or more ofthe peaks set forth in Table 2. In another embodiment, Form A ofCompound A has one or more characteristic X-ray powder diffraction peaksat a two-theta angle of approximately 8.3, 8.8, 12.0, 13.2, 13.9, 14.4,14.8, 16.5, 17.7, 18.2, 19.3, 19.5, 19.6, 21.0, 21.2, 21.7, 22.5, 24.1,24.7, 25.0, 25.3, 26.5, 26.7, 28.3, 29.3, 29.5, 29.8, 30.5, 32.1, 33.3,34.2 or 34.6 degrees. In a specific embodiment, Form A of Compound A hasone, two, three, four, five, six, seven or eight characteristic X-raypowder diffraction peaks at a two-theta angle of approximately 8.3, 8.8,13.2, 16.5, 17.7, 18.2, 21.7 or 26.5 degrees. In another embodiment,Form A of Compound A has one, two, three or four characteristic X-raypowder diffraction peaks at a two-theta angle of approximately 8.3,13.2, 18.2 or 21.7 degrees. In a particular embodiment, Form A ofCompound A has one or more characteristic X-ray powder diffraction peaksat a two-theta angle of approximately 8.0, 9.0, 12.0, 13.0, 16.5, 17.5,18.2, 21.5, 22.5, 25.0 or 26.5 degrees. In a specific embodiment, Form Aof Compound A has one, two, three, four, five, six, seven or eightcharacteristic X-ray powder diffraction peaks at a two-theta angle ofapproximately 8.0, 9.0, 13.0, 16.5, 17.5, 18.2, 21.5 or 26.5 degrees. Inanother embodiment, Form A of Compound A has one, two, three or fourcharacteristic X-ray powder diffraction peaks at a two-theta angle ofapproximately 8.0, 13.0, 18.2 or 21.5 degrees. In another embodiment,Form A of Compound A has one, two, three or four characteristic X-raypowder diffraction peaks at a two-theta angle of approximately 13.0,16.5, 18.2 or 21.5 degrees.

In another embodiment, Form A of Compound A has a thermogravimetricthermogram substantially as shown in FIG. 3. In certain embodiments,Form A of Compound A shows less than about 10%, less than about 5%, lessthan about 3%, less than about 2%, less than about 1%, less than about0.5%, less than about 0.2%, less than about 0.1%, less than about 0.05%,or less than about 0.03%, e.g., about 0.024%, weight loss between about25° C. to about 100° C. in a thermogravimetric thermogram. In certainembodiments, Form A of Compound A shows less than about 0.1% weight lossbetween about 25° C. to about 100° C. in a thermogravimetric thermogram.In certain embodiments, Form A of Compound A shows about 0.025% weightloss between about 25° C. to about 100° C. in a thermogravimetricthermogram. In certain embodiments, Form A of Compound A shows no weightloss until degradation at about 260° C. in a thermogravimetricthermogram. In certain embodiments, Form A of Compound A is anhydrous.In certain embodiments, Form A of Compound A is unsolvated.

In yet another embodiment, Form A of Compound A has a differentialscanning calorimetric (DSC) thermogram substantially as shown in FIG. 4.In certain embodiments, Form A of Compound A has an endotherm with apeak temperature of about 201° C. in a DSC thermogram. In certainembodiments, Form A of Compound A has an endotherm with an onsettemperature of about 197° C. in a DSC thermogram. In certainembodiments, Form A of Compound A has an endotherm with a peaktemperature of about 199° C. and an onset temperature of about 197° C.in a DSC thermogram. In one embodiment, Form A of Compound A has amelting temperature of about 197-199° C. In certain embodiment, Form Aof Compound A has a melting temperature of about 199° C. In oneembodiment, Form A of Compound A has an endotherm of about 195° C. in aDSC thermogram.

In yet another embodiment, Form A of Compound A is non-hygroscopic,e.g., exhibits a mass gain of less than about 0.1% w/w of when subjectedto an increase in humidity from about 0% to about 80% relative humidity(RH). In another embodiment, Form A of Compound exhibits a mass gain ofabout 0.5% w/w of when subjected to an increase in humidity from about80% to about 90% relative humidity. In certain embodiments, Form A ofCompound A exhibits no greater than about 2% w/w, no greater than about1% w/w, no greater than about 0.6% w/w, no greater than about 0.4% w/w,no greater than about 0.2% w/w, or no greater than about 0.1% w/w weightgain in response to an increase in humidity from about 0% to about 95%relative humidity at about 25° C. In certain embodiments, Form A ofCompound A exhibits about 0.3% w/w weight gain in response to anincrease in humidity from about 0% to about 95% relative humidity atabout 25° C. In certain embodiments, Form A of Compound A exhibits nogreater than about 2% w/w, no greater than about 1% w/w, no greater thanabout 0.6% w/w, no greater than about 0.4% w/w, no greater than about0.2% w/w, or no greater than about 0.1% w/w weight gain in response toan increase in humidity from about 0% to about 50% relative humidity atabout 25° C. In certain embodiments, Form A of Compound A exhibits about0.1% w/w weight gain in response to an increase in humidity from about0% to about 50% relative humidity at about 25° C.

In one embodiment, provided herein is Form B of Compound A. In oneembodiment, Form B of Compound A has an X-ray powder diffraction patternsubstantially as shown in FIG. 9. In another embodiment, Form B ofCompound A has one or more characteristic X-ray powder diffraction peaksat a two-theta angle of approximately 6.0, 7.0, 8.0, 10.0, 12.0, 14.0,17.0, 18.0, 20.0, 20.5, 22.5, or 24.5 degrees. In a specific embodiment,Form B of Compound A has one, two, three, four, five, six, or sevencharacteristic X-ray powder diffraction peaks at a two-theta angle ofapproximately 6.0, 7.0, 8.0, 10.0, 12.0, 14.0, 17.0, 18.0, 20.0, 20.5,22.5, or 24.5 degrees. In another embodiment, Form B of Compound A hasone, two, three or four characteristic X-ray powder diffraction peaks ata two-theta angle of approximately 6.0, 7.0, 8.0, 10.0, 12.0, 14.0,17.0, 18.0, 20.0, 20.5, 22.5, or 24.5 degrees.

In certain embodiments, Form B of Compound A shows less than about 10%or less than about 7%, e.g., about 6.4%, weight loss and an onsettemperature of about 50° C. in a thermogravimetric thermogram. Incertain embodiments, Form B of Compound A is a hydrate.

In yet another embodiment, Form B of Compound A has a differentialscanning calorimetric (DSC) thermogram substantially as shown in FIG.10. In certain embodiments, Form B of Compound A has an endotherm with apeak temperature of about 111.3° C., and an exotherm with a peaktemperature of about 164.9° C. in a DSC thermogram. In certainembodiments, Form B of Compound A has an endotherm with a peaktemperature of about 202° C.

In one embodiment, provided herein is Form C of Compound A. In oneembodiment, Form C of Compound A has an X-ray powder diffraction patternsubstantially as shown in FIG. 11. In another embodiment, Form C ofCompound A has one or more characteristic X-ray powder diffraction peaksat a two-theta angle of approximately 6.5, 9.0, 10.0, 14.5, 16.5, 19.0,23.0, or 23.5 degrees. In a specific embodiment, Form C of Compound Ahas one, two, three, four, five, six, or seven characteristic X-raypowder diffraction peaks at a two-theta angle of approximately 6.5, 9.0,10.0, 14.5, 16.5, 19.0, 23.0, or 23.5 degrees. In another embodiment,Form C of Compound A has one, two, three or four characteristic X-raypowder diffraction peaks at a two-theta angle of approximately 6.5, 9.0,10.0, 14.5, 16.5, 19.0, 23.0, or 23.5 degrees. In a particularembodiment, Form C of Compound A has one or more characteristic X-raypowder diffraction peaks at a two-theta angle of approximately 6.5, 9.0,10.0, 14.5, 16.5, 19.0, 23.0, or 23.5 degrees.

In certain embodiments, Form C of Compound A is anhydrous.

In yet another embodiment, Form C of Compound A has a differentialscanning calorimetric (DSC) thermogram substantially as shown in FIG.12. In certain embodiments, Form C of Compound A has an endotherm andexotherm of about 160° C. and an endotherm of about 200° C. in a DSCthermogram. In certain embodiments, Form C of Compound A has anendotherm of about 162° C. and an endotherm of about 200° C. in a DSCthermogram.

In one embodiment, provided herein is Form D of Compound A. In oneembodiment, Form D of Compound A has an X-ray powder diffraction patternsubstantially as shown in FIG. 14. In another embodiment, Form D ofCompound A has one or more characteristic X-ray powder diffraction peaksat a two-theta angle of approximately 6.0, 8.0, 9.0, 10.0, 12.5, 14.5,16.5, 18.0, 19.0, 19.5, 20.5, 22.5, 23.5, or 27.5 degrees. In a specificembodiment, Form D of Compound A has one, two, three, four, five, six,seven, eight, nine, ten, eleven, or twelve characteristic X-ray powderdiffraction peaks at a two-theta angle of approximately 6.0, 7.5, 8.0,9.0, 10.0, 12.5, 14.5, 16.5, 19.0, 19.5, 20.5, or 23.0 degrees. Inanother embodiment, Form D of Compound A has one, two, three or fourcharacteristic X-ray powder diffraction peaks at a two-theta angle ofapproximately 6.0, 7.5, 8.0, 9.0, 10.0, 12.5, 14.5, 16.5, 19.0, 19.5,20.5, or 23.0 degrees. In a particular embodiment, Form D of Compound Ahas one or more characteristic X-ray powder diffraction peaks at atwo-theta angle of approximately 6.0, 7.5, 8.0, 9.0, 10.0, 12.5, 14.5,16.5, 19.0, 19.5, 20.5, or 23.0 degrees.

In certain embodiments, Form D of Compound A shows less than about 10%or less than about 8%, e.g., about 7.4%, weight loss and an onsettemperature of about 80° C. in a thermogravimetric thermogram. Incertain embodiments, Form D of Compound A is a solvate.

In yet another embodiment, Form D of Compound A has a differentialscanning calorimetric (DSC) thermogram substantially as shown in FIG.13. In certain embodiments, Form D of Compound A has an endotherm with apeak temperature of about 98.3° C., and an endotherm with a peaktemperature of about 159.3° C. in a DSC thermogram. In certainembodiments, Form D of Compound A has an endotherm with a peaktemperature of about 200.6° C.

In one embodiment, Form E is a p-xylene solvated form of Compound A. Inone embodiment, Form E is a p-xylene hemi-solvated form of Compound A.In another embodiment, Form E of Compound A is crystalline.

In one embodiment, a single-crystal X-ray diffraction analysis isemployed to determine the crystal structure of Form E of Compound A.Table 4 and Table 5 present a summary of the crystallographic data fromthe crystal-structure determination. In one embodiment, Form E ofCompound A has a crystal packing pattern substantially as shown in FIG.16. In one embodiment, Form E is a p-xylene hemi-solvated formcrystallizing in a monoclinic crystal system. In one embodiment, Form Eis a p-xylene hemi-solvated form crystallizing in a monoclinic crystalsystem with a P2₁/c space group.

In certain embodiments, a solid form provided herein (e.g., Form E) issubstantially crystalline, as indicated by, e.g., X-ray powderdiffraction measurements. In one embodiment, Form E of Compound A has anX-ray powder diffraction pattern substantially as shown in FIG. 15 (thesecond pattern from the bottom). In one embodiment, Form E of Compound Ahas one or more characteristic X-ray powder diffraction peaks at atwo-theta angle of approximately 7.46, 8.94, 11.7, 13.7, 17.26, 18.22,18.78, 20.94, 22.38, 23.06 or 24.62 degrees as depicted in FIG. 15 (thesecond pattern from the bottom). In another embodiment, Form E ofCompound A has one, two, three, four five, six, seven or eightcharacteristic X-ray powder diffraction peaks at a two-theta angle ofapproximately 7.46, 8.94, 13.7, 18.22, 18.78, 22.38, 23.06 or 24.62degrees. In another embodiment, Form E of Compound A has one, two, threeor four characteristic X-ray powder diffraction peaks at a two-thetaangle of approximately 7.46, 13.7, 18.22 or 23.06 degrees. In anotherembodiment, Form E of Compound A has one, two, three, four, five, six,seven, eight, nine, ten or eleven characteristic X-ray powderdiffraction peaks as set forth in Table 3.

In certain embodiments, Form E of Compound A has digital imagessubstantially as shown in FIG. 17A and FIG. 17B.

In one embodiment, provided herein is a crystalline form of Compound Ahaving a thermogravimetric (TGA) thermograph corresponding substantiallyto the representative TGA thermogram as depicted in FIG. 19. In certainembodiments, the crystalline form exhibits a TGA thermogram comprising atotal mass loss of approximately 11.0%-11.5% (e.g., 11.0% or 11.2%) ofthe total mass of the sample between approximately 50° C. andapproximately 200° C. when heated from approximately 25° C. toapproximately 300° C. Thus, in certain embodiments, the crystalline formloses about 11.0% of its total mass when heated from about ambienttemperature to about 300° C. In certain embodiments, the crystallineform contains 0.5 molar equivalents of solvent in the crystal latticecorresponding to approximately 0.5 mole of p-xylene per mole of CompoundA. The theoretical p-xylene content of a p-xylene hemi-solvate ofCompound A is 10.5% by weight, matching the TGA weight loss observed. Incertain embodiments, the crystalline form is a p-xylene hemi-solvate ofCompound A.

In one embodiment, provided herein is a crystalline form of Compound Ahaving a single differential thermal analysis (SDTA) thermogram asdepicted in FIG. 18 comprising an endothermic event between about 90° C.and about 125° C. with a maximum at about 106-110° C., when heated fromapproximately 25° C. to approximately 300° C.

In one embodiment, provided herein is a crystalline form of Compound Ahaving a SDTA thermogram comprising an endothermic event as depicted inFIG. 17 with a maximum at about 193° C. when heated from approximately25° C. to approximately 300° C.

In still another embodiment, Form A of Compound A is substantially pure.In certain embodiments, the substantially pure Form A of Compound A issubstantially free of other solid forms, e.g., amorphous form. Incertain embodiments, the purity of the substantially pure Form A ofCompound A is no less than about 95% pure, no less than about 96% pure,no less than about 97% pure, no less than about 98% pure, no less thanabout 98.5% pure, no less than about 99% pure, no less than about 99.5%pure, or no less than about 99.8% pure.

In still another embodiment, Form B of Compound A is substantially pure.In certain embodiments, the substantially pure Form B of Compound A issubstantially free of other solid forms, e.g., amorphous form. Incertain embodiments, the purity of the substantially pure Form B ofCompound A is no less than about 95% pure, no less than about 96% pure,no less than about 97% pure, no less than about 98% pure, no less thanabout 98.5% pure, no less than about 99% pure, no less than about 99.5%pure, or no less than about 99.8% pure.

In still another embodiment, Form C of Compound A is substantially pure.In certain embodiments, the substantially pure Form C of Compound A issubstantially free of other solid forms, e.g., amorphous form. Incertain embodiments, the purity of the substantially pure Form C ofCompound A is no less than about 95% pure, no less than about 96% pure,no less than about 97% pure, no less than about 98% pure, no less thanabout 98.5% pure, no less than about 99% pure, no less than about 99.5%pure, or no less than about 99.8% pure.

In still another embodiment, Form D of Compound A is substantially pure.In certain embodiments, the substantially pure Form D of Compound A issubstantially free of other solid forms, e.g., amorphous form. Incertain embodiments, the purity of the substantially pure Form D ofCompound A is no less than about 95% pure, no less than about 96% pure,no less than about 97% pure, no less than about 98% pure, no less thanabout 98.5% pure, no less than about 99% pure, no less than about 99.5%pure, or no less than about 99.8% pure.

In still another embodiment, Form E of Compound A is substantially pure.In certain embodiments, the substantially pure Form E of Compound A issubstantially free of other solid forms, e.g., amorphous form. Incertain embodiments, the purity of the substantially pure Form E ofCompound A is no less than about 95% pure, no less than about 96% pure,no less than about 97% pure, no less than about 98% pure, no less thanabout 98.5% pure, no less than about 99% pure, no less than about 99.5%pure, or no less than about 99.8% pure.

In one embodiment, provided herein is a pinacol co-crystal of CompoundA. In one embodiment, the pinacol co-crystal of Compound A has an X-raypowder diffraction pattern substantially as shown in FIG. 7. In anotherembodiment, the pinacol co-crystal of Compound A has one or morecharacteristic X-ray powder diffraction peaks at a two-theta angle ofapproximately 5.0, 6.0, 12.5, 14.0, 15.0, 15.5, 17.5, 18.5, and 22.5degrees. In a specific embodiment, the pinacol co-crystal of Compound Ahas one, two, three, four, or five characteristic X-ray powderdiffraction peaks at a two-theta angle of approximately 5.0, 6.0, 12.5,14.0, 15.0, 15.5, 17.5, 18.5, and 22.5 degrees. In another embodiment,the pinacol co-crystal of Compound A has one, two, three or fourcharacteristic X-ray powder diffraction peaks at a two-theta angle ofapproximately 5.0, 6.0, 12.5, 14.0, 15.0, 15.5, 17.5, 18.5, and 22.5degrees.

In yet another embodiment, the pinacol co-crystal of Compound A has adifferential scanning calorimetric (DSC) thermogram substantially asshown in FIG. 6. In certain embodiments, the pinacol co-crystal ofCompound A has an endotherm with a peak temperature of about 119° C. ina DSC thermogram. In certain embodiments, the pinacol co-crystal ofCompound A has an endotherm with an onset temperature of about 115° C.in a DSC thermogram. In certain embodiments, the pinacol co-crystal ofCompound A has an endotherm with a peak temperature of about 119° C. andan onset temperature of about 115° C. in a DSC thermogram. In anotherembodiment, the pinacol co-crystal of Compound A is comprised of about20% by weight of pinacol.

In still another embodiment, the pinacol co-crystal of Compound A issubstantially pure. In certain embodiments, the substantially purepinacol co-crystal of Compound A is substantially free of other solidforms, e.g., amorphous form. In certain embodiments, the purity of thesubstantially pure pinacol co-crystal of Compound A is no less thanabout 95% pure, no less than about 96% pure, no less than about 97%pure, no less than about 98% pure, no less than about 98.5% pure, noless than about 99% pure, no less than about 99.5% pure, or no less thanabout 99.8% pure.

The solid forms of Compound A provided herein (for example, Forms A, B,C, D or E) can be prepared by the methods described herein.

In certain embodiments, Form A of Compound A can be prepared by solventevaporation of a solution or slurry of Compound A in toluene, MTBE(methyl tert-butyl ether), DIPE (diisopropyl ether), THF(tetrahydrofuran), DME (dimethoxyethane), IPAc (isopropyl acetate),EtOAc (ethyl acetate), MIBK (methyl isobutyl ketone), acetone, IPA(isopropyl alcohol), ethanol, ACN (acetonitrile), nitromethane orIPA:water (for example, 95:5).

In certain embodiments, Form A of Compound A can be prepared bysubjecting a solution or slurry of Compound A in toluene, MTBE (methyltert-butyl ether), DIPE (diisopropyl ether), THF (tetrahydrofuran), DME(dimethoxyethane), IPAc (isopropyl acetate), EtOAc (ethyl acetate), MIBK(methyl isobutyl ketone), acetone, IPA (isopropyl alcohol), ethanol, ACN(acetonitrile), nitromethane or IPA:water (95:5) to cycles of heating toabout 50° C. and cooling to room temperature, followed by solventevaporation.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving amorphous7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-onein toluene, MTBE (methyl tert-butyl ether), DIPE (diisopropyl ether),THF (tetrahydrofuran), DME (dimethoxyethane), IPAc (isopropyl acetate),EtOAc (ethyl acetate), MIBK (methyl isobutyl ketone), acetone, IPA(isopropyl alcohol), ethanol, ACN (acetonitrile), nitromethane, orIPA:water (95:5) and allowing the resulting solution to evaporate atroom temperature.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT (butylated hydroxytoluene), IPA and water, heatingand then cooling to room temperature. In some embodiments, the methodsfurther comprise collection by filtration, washing with IPA and waterand drying.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT and MeOAc (methyl acetate), heating, cooling to roomtemperature, distilling under vacuum and contacting with n-heptane. Incertain embodiments, the methods further comprise collection byfiltration and washing with MeOAc and n-heptane and drying. In certainembodiments, this method further comprises adding a small amount of FormA in MeOAc to the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein BHT and MeOAc. In some embodiments, the methods further comprisefiltration of the hot BHT and MeOAc solution.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT and MeOAc (methyl acetate), heating, filtering,cooling, distilling under vacuum and contacting with n-heptane. Incertain embodiments, the methods further comprise collection byfiltration and washing with MeOAc and n-heptane and drying. In certainembodiments, this method further comprises adding a small amount of FormA in MeOAc to the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein BHT and MeOAc. In some embodiments, the methods further comprisefiltration of the hot BHT and MeOAc solution.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-oneand7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onepinacol cocrystal in IPA. In some embodiments, the methods furthercomprise collection by filtration and drying, such as under reducedpressure.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT, THF and water, contacting with IPAc, heating,distilling, cooling, contacting with IPAc, heating, distilling, and thencooling to room temperature. In some embodiments, the methods furthercomprise collection by filtration, washing with IPAc, and drying. Insome embodiment, the method additionally comprises treating the BHT, THFand water solution with activated carbon. In certain embodiments, themethod further comprises filtration of the BHT, THE and water solution.In some embodiments, the methods comprise a first distillation step atatmospheric pressure at constant volume (by addition of IPAc) and thencooling. In certain embodiments, this method further comprises adding asmall amount of Form A in IPAc after the first distillation. In someembodiments, the method further comprises a second distillation atatmospheric pressure at constant volume ((by addition of IPAc) and thencooling.

In certain embodiments, provided herein are methods for making Form A of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT, THF and water, contacting with IPAc, heating,distilling under reduced pressure, contacting with IPAc, and thencooling to room temperature. In some embodiments, the methods furthercomprise collection by filtration, washing with IPAc, and drying. Insome embodiment, the method additionally comprises treating the BHT, THFand water solution with activated carbon. In certain embodiments, themethod further comprises filtration of the BHT, THF and water solution.In some embodiments, the methods comprise a first distillation step atreduced pressure at constant temperature, with addition of IPAc. Incertain embodiments, this method further comprises adding a small amountof Form A in IPAc after the first distillation. In some embodiments, themethod further comprises a second distillation under vacuum, withaddition of IPAc, and then cooling to room temperature.

In certain embodiments, provided herein are methods for making Form B of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT, IPA and water, heating the mixture and addingwater, cooling the mixture, collection by filtration, washing with IPAand water, and drying. In certain embodiments, this method furthercomprises adding a small amount of Form B in water to the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein BHT, IPA and water.

In certain embodiments, provided herein are methods for making Form C of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT, MeOH, distilling to remove MeOH, furtherdistillation with IPA, cooling the mixture, collection by filtration,washing with IPA and drying.

In certain embodiments, provided herein are methods for making Form D of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising dissolving7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onein a mixture of BHT in MeOH, heating, then cooling with stirring,collection by filtration, washing and drying.

In certain embodiments, Form E is obtained by crystallization fromcertain solvent systems, for example, solvent systems comprising one ormore of the following solvents or solvent combinations: p-xylene/acetone(e.g., 50/50), p-xylene/MTBE (e.g., 50/50) and p-xylene.

In certain embodiments, provided herein are methods for making Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising obtaining a slurry of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-onein a solvent, heating the slurry to a first temperature (e.g., about 50°C. to about 70° C.), filtering the slurry to yield a solution, coolingdown the solution to a second temperature (e.g., about 15° C. to about35° C.) to yield solids, and collecting the solids. In certainembodiments, provided herein are methods for making Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising obtaining a slurry of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-onein p-xylene/acetone (50/50), heating the slurry to about 60° C.,filtering the slurry to yield a solution, cooling down the solution toabout 25° C. to yield solids, and collecting the solids.

In certain embodiments, provided herein are methods for making Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising mixing7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-onewith a solvent, filtering the mixture to yield a solution if7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-onedoes not dissolve completely, and evaporating the solution under certainair pressure to yield a solid. In certain embodiments, provided hereinare methods for making Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising mixing7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-onewith p-xylene/MTBE (50/50), filtering the mixture to yield a solution if7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-onedoes not dissolve completely, and evaporating the solution under 200mbar air pressure to yield a solid.

In certain embodiments, provided herein are methods for making Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising obtaining a slurry of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-onein a solvent, stirring the slurry, collecting solids from the slurry byfiltration (e.g., centrifuge filtration) and optionally washing (e.g.,washing with the solvent) and drying. In certain embodiments, providedherein are methods for making Form E of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,comprising obtaining a slurry of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-onein p-xylene, stirring the slurry, collecting solids from the slurry bycentrifuge filtration and optionally washing with p-xylene and drying.

In certain embodiments, provided herein are methods for making a pinacolco-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,comprising mixing7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onewith pinacol in solution (for example THF and toluene), heating untilsolids are dissolved, distilling said solution and seeding with apinacol co-crystal of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.IN some embodiments, the methods further comprise collection byfiltration, washing with THF/toluene and drying.

5.3 Process of Preparation of Compound A

In certain embodiments, provided herein are methods for preparingCompound A, comprising: (1) contactingethyl-2-(3,5-dibromopyrazin-2-ylamino)acetate withtrans-4-methoxycyclohexylamine hydrochloride and 1-methyl-2-pyrrolidineand adding DIPEA to produce ethyl2-((5-bromo-3-(((trans)-4-methoxycyclohexyl)amino)pyrazin-2-yl)amino)acetate;(2) contacting ethyl2-((5-bromo-3-(((trans)-4-methoxycyclohexyl)amino)pyrazin-2-yl)amino)acetatewith an acid (such as a phosphoric acid solution) to produce7-bromo-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one;and (3) contacting7-bromo-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-onewith2-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)propan-2-oland PdCl₂(Amphos)₂.

Provided herein are methods of preparing Compound A

the method comprising contacting a compound of Formula b

with a compound of formula c

in a solvent (e.g., THF), in the presence of a base (e.g., K₂CO₃) and apalladium catalyst (e.g., PdCl₂(Amphos)₂), wherein said contactingoccurs under conditions suitable to provide Compound A. In someembodiments, the contacting occurs at elevated temperature (e.g.,reflux). In certain embodiments, the palladium catalyst ischloro(2-dicyclohexylphosphino-2′, 6′-di-i-propoxy-1,1′-biphenyl)[2-(2-aminoethylphenyl)]palladium(II),chloro(2-dicyclohexylphosphino-2′,6′-di-i-propoxy-1,1′-biphenyl)(2-amino-1,1′-biphenyl-2-yl)palladium(II),chloro(2-dicyclohexylphosphino-2′,6′-dimethoxy-1,1′-biphenyl)[2-(2-aminoethylphenyl)]palladium(II)dichloromethane adduct,chloro(2-dicyclohexylphosphino-2′,6′-dimethoxy-1,1′-biphenyl)(2′-amino-1,1′-biphenyl-2-yl)palladium(II),chloro(2-dicyclohexylphosphino-2′,4′,6′-tri-i-propyl-1,1′-biphenyl)[2-(2-aminoethyl)phenyl]palladium(II),chloro(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II),chloro[2-(dicyclohexylphosphino)-3,6-dimethoxy-2′-4′-6′-tri-i-propyl-1,1′-biphenyl][2-(2-aminoethyl)phenyl]palladium(II),[1,1′-bis(di-tert-butylphosphino)ferrocene]dichloropalladium,[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium,bis(triphenylphosphine)palladium(II) chloride complex,tetrakis(triphenylphosphine)palladium(0),dichlorobis(tricyclohexylphoshine)palladium(II), palladium(II) chloridewith 2′-(dicyclohexylphosphino)acetophenone ethylene ketal ligand,palladium (II) chloride with1,2,3,4,5-pentaphenyl-1′-(di-tert-butylphosphino)ferrocene ligand,palladium (II) chloride with2-dicyclohexylphosphino-2′-(N,N-dimethylamino)biphenyl ligand, palladium(II) chloride with 4,5-bis(diphenylphosphino)-9,9-dimethylxantheneligand or palladium (II) chloride with2-dicyclohexylphosphino-2′-methylbiphenyl ligand.

In some such embodiments, the methods further comprise preparing acompound of formula b

the method comprising contacting a compound of formula d

with an acid (e.g., phosphoric acid), wherein said contacting occursunder conditions suitable to provide a compound of formula b. In someembodiments, the contacting occurs at elevated temperature (e.g., 80°C.).

In some such embodiments, the methods further comprise preparing acompound of formula d

the method comprising contacting a compound of formula e

with trans-4-methoxycyclohexylamine hydrochloride, in the presence of abase (e.g., DIPEA), in a solvent (e.g., NMP), wherein said contactingoccurs under conditions suitable to provide a compound of formula b.

In some embodiments, the contacting occurs at elevated temperature(e.g., 125-130° C.).

Isotopologues of Compound A and metabolites thereof can be prepared bythe methods provided herein.

In one embodiment, provided herein are processes for preparing acompound having the formula:

the method comprising contacting

in the presence of a palladium catalyst (e.g., PdCl₂(Amphos)₂) and abase (e.g., K₂CO₃) in a solvent (e.g., THF, optionally with water),wherein said contacting occurs under conditions suitable to produce

In some embodiments, the contacting occurs at elevated temperature(e.g., 73° C.).

In one embodiment, provided herein are processes for preparing acompound having the formula:

the method comprising contacting

in the presence of a palladium catalyst (e.g., PdCl₂(Amphos)₂) and abase (e.g., K₂CO₃) in a solvent (e.g., THF, optionally with water),wherein said contacting occurs under conditions suitable to produce

In some such embodiments, the contacting occurs at elevated temperature(e.g., 73° C.). In some such embodiments, the method further comprisesaddition of EtOAc, and isolation of crude ¹⁴C-compound A using EtOAc,DCM, methanol, and silica gel and drying. In some such embodiments,crude ¹⁴C-compound A is dissolved in BHT and ACN and isolated usingEtOAc.

In some embodiments, the methods further comprise contacting

with an acid (e.g., HCl) in a solvent (e.g., 1,4-dioxane), wherein saidcontacting occurs under conditions suitable to produce

In some embodiments, the methods further comprise contacting

in the presence of a palladium catalyst (e.g., PdCl₂(dppf)-DCM complex)and a base (e.g., KOAc) in a solvent (e.g., 1,4-dioxane), wherein saidcontacting occurs under conditions suitable to produce

In some embodiments, the methods further comprise contacting

with TMSCl in the presence of a base (e.g., TEA) in a solvent (e.g.,DCM), wherein said contacting occurs under conditions suitable toproduce

In some embodiments, the methods further comprise contacting

in the presence of a base (e.g., butyl lithium) in a solvent (e.g., DCM)wherein said contacting occurs under conditions suitable to produce

Further provided herein are processes for preparing a compound havingthe formula:

the method comprising contacting

with an acid (aqueous HCl) in a solvent (e.g., ACN) wherein saidcontacting occurs under conditions suitable to produce

In some embodiments, the methods further comprise contacting

in the presence of a palladium catalyst (e.g., PdCl₂(Amphos)₂) and abase (e.g., K₂CO₃) in a solvent (e.g., IPA, optionally in the presenceof water), wherein said contacting occurs under conditions suitable toproduce

In some such embodiments, the contacting occurs at eleveatedtemperatures (e.g., 69-71° C.).

In some embodiments, the methods further comprise contacting

in the presence of a palladium catalyst (e.g., PdCl₂(dppf)-DCM complex)and a base (e.g., K₂CO₃) in a solvent (e.g 1,4-dioxane), wherein saidcontacting occurs under conditions suitable to produce

In some such embodiments, the contacting occurs at elevated temperature(e.g., 90-95° C.).

In some embodiments, the methods further comprise contacting

with TMSCl in the presence of a base (e.g., TEA, optionally in thepresence of DMAP) in a solvent (e.g., in DCM), wherein said contactingoccurs under conditions suitable to produce

In some such embodiments, the contacting occurs at low temperature(e.g., 0-5° C.).

In some embodiments, the methods further comprise contacting

in the presence of a base (e.g., n-butyllithium) in a solvent (e.g.,DCM), wherein said contacting occurs under conditions suitable toproduce

In some such embodiments, the contacting occurs at low temperature(e.g., −78° C.).

In some embodiments, the methods further comprise contacting

with a base (e.g., potassium tert-butoxide) in a solvent (e.g THF),wherein said contacting occurs under conditions suitable to produce

In some other embodiments, the methods further comprise contacting

with and acid (e.g. phosphoric acid) in a solvent (e.g water), whereinsaid contacting occurs under conditions suitable to produce

In some such embodiments, the contacting occurs at elevated temperature(e.g., 80° C.).

In some embodiments, the methods further comprise contacting

in the presence of a base (e.g., DIPEA) in a solvent (e.g., NMP),wherein said contacting occurs under conditions suitable to produce

In some such embodiments, the contacting occurs at elevated temperature(e.g., 124-129° C.).

In some embodiments, the methods further comprise contacting

in the presence of a base (e.g., K₂CO₃) in a solvent e.g acetone),optionally in the presence of tetrabutylammonium hydrogensulfate,wherein said contacting occurs under conditions suitable to produce

In one embodiment, provided herein are methods of preparing a compoundhaving the formula:

the methods comprising contacting

in the presence of a palladium catalyst (e.g., PdCl₂(Amphos)₂) and abase (e.g., K₂CO₃) in a solvent (e.g., THF, optionally with water),wherein said contacting occurs under conditions suitable to produce

In some such embodiments, the contacting occurs at elevated temperatures(e.g., reflux).

In some embodiments, the methods further comprise contacting

with an acid (e.g., HCl) in a solvent (e.g., 1,4-dioxane), wherein saidcontacting occurs under conditions suitable to produce

In some embodiments, the methods further comprise contacting

in the presence of a palladium catalyst (e.g., PdCl₂(dppf)-DCM complex)and a base (e.g., K₂CO₃) in a solvent (e,g. 1,4-dioxane), wherein saidcontacting occurs under conditions suitable to produce

In some embodiments, the contacting occurs at elevated temperature (e.g.reflux).

In some embodiments, the methods further comprise contacting

with TMSCl in the presence of a base (e.g., TEA, optionally in thepresence of DMAP) in a solvent (e.g., DCM), wherein said contactingoccurs under conditions suitable to produce

In some embodiments, the methods further comprise contacting

in the presence of a base (e.g n-butyllithium) in a solvent (e.g., DCM),wherein said contacting occurs under conditions suitable to produce

In some embodiments, the contacting occurs at low temperature (e.g.,−78° to −72° C.).

In some embodiments, the methods further comprise contacting

with an acid (e.g., aqueous phosphoric acid), wherein said contactingoccurs under conditions suitable to produce

In some embodiments, the contacting occurs at elevated temperature(e.g., 75-80° C.).

In some embodiments, the methods further comprise contacting

in the presence of a base (e.g., DIPEA) in a solvent (e.g., NMP),wherein said contacting occurs under conditions suitable to produce

In some embodiments, the contacting occurs at elevated temperature(e.g., reflux).

In some embodiments, the methods further comprise contacting

in the presence of a base (e.g., K₂CO₃) in a solvent (e.g., acetone),optionally in the presence of tetrabutylammonium hydrogensulfate,wherein said contacting occurs under conditions suitable to produce

In some embodiments, the contacting occurs at elevated temperature(e.g., reflux).

In one embodiment, the compound having the formula:

is recrystallized from a mixture of 2-propanol and water in the presenceof 2,6-di-tert-butyl-4-methylphenol.

In one embodiment, provided herein are methods of preparing a compoundhaving the formula:

the methods comprising contacting

in the presence of a palladium catalyst (e.g., PdCl₂(Amphos)₂) and abase (e.g., K₂CO₃) in a solvent (e.g., THF, optionally with water),wherein said contacting occurs under conditions suitable to produce

In some such embodiments, the contacting occurs at elevated temperatures(e.g., reflux).

In some embodiments, the methods further comprise contacting

with an acid (e.g., HCl) in a solvent (e.g., 1,4-dioxane), wherein saidcontacting occurs under conditions suitable to produce

In some embodiments, the methods further comprise contacting

in the presence of a palladium catalyst (e.g., PdCl₂(dppf)-DCM complex)and a base (e.g., K₂CO₃) in a solvent (e,g. 1,4-dioxane), wherein saidcontacting occurs under conditions suitable to produce

In some embodiments, the contacting occurs at elevated temperature (e.g.reflux).

In some embodiments, the methods further comprise contacting

with TMSCl in the presence of a base (e.g., n-butyllithium) in a solvent(e.g., d₆-acetone), wherein said contacting occurs under conditionssuitable to produce

In some embodiments, the methods further comprise contacting

with an acid (e.g., aqueous phosphoric acid), wherein said contactingoccurs under conditions suitable to produce

In some embodiments, the contacting occurs at elevated temperature(e.g., 75-80° C.).

In some embodiments, the methods further comprise contacting

in the presence of a base (e.g., DIPEA) in a solvent (e.g., NMP),wherein said contacting occurs under conditions suitable to produce

In some embodiments, the contacting occurs at elevated temperature(e.g., reflux).

In some embodiments, the methods further comprise contacting

in the presence of a base (e.g., K₂CO₃) in a solvent (e.g., acetone),optionally in the presence of tetrabutylammonium hydrogensulfate,wherein said contacting occurs under conditions suitable to produce

In some embodiments, the contacting occurs at elevated temperature(e.g., reflux).

In one embodiment, provided herein are methods of preparing a compoundhaving the formula:

the methods comprising contacting

in the presence of a palladium catalyst (e.g., PdCl₂(Amphos)₂) and abase (e.g., K₂CO₃) in a solvent (e.g., THF, optionally with water),wherein said contacting occurs under conditions suitable to produce

In some such embodiments, the contacting occurs at elevated temperatures(e.g., reflux).

In some embodiments, the methods further comprise contacting

with an acid (e.g., HCl) in a solvent (e.g., 1,4-dioxane), wherein saidcontacting occurs under conditions suitable to produce

In some embodiments, the methods further comprise contacting

in the presence of a palladium catalyst (e.g., PdCl₂(dppf)-DCM complex)and a base (e.g., K₂CO₃) in a solvent (e,g. 1,4-dioxane), wherein saidcontacting occurs under conditions suitable to produce

In some embodiments, the contacting occurs at elevated temperature(e.g., reflux).

In some embodiments, the methods further comprise contacting

with TMSCl in the presence of a base (e.g., n-butyllithium) in a solvent(e.g., d⁶-acetone), wherein said contacting occurs under conditionssuitable to produce

In some embodiments, the methods further comprise contacting

with an acid (e.g., aqueous phosphoric acid), wherein said contactingoccurs under conditions suitable to produce

In some embodiments, the contacting occurs at elevated temperature(e.g., 75-80° C.).

In some embodiments, the methods further comprise contacting

in the presence of a base (e.g., DIPEA) in a solvent (e.g., NMP),wherein said contacting occurs under conditions suitable to produce

In some embodiments, the contacting occurs at elevated temperature(e.g., reflux).

In some embodiments, the methods further comprise contacting

in the presence of a base (e.g., K₂CO₃) in a solvent (e.g., acetone),optionally in the presence of tetrabutylammonium hydrogensulfate,wherein said contacting occurs under conditions suitable to produce

In some embodiments, the contacting occurs at elevated temperature(e.g., reflux).

In one embodiment, the compound having the formula:

In one embodiment, provided herein are methods of preparing a compoundhaving the formula:

the methods comprising contacting

with a base and CD₃I to produce

further contacting with a base and ROD/D₂O, wherein said contactingoccurs under conditions suitable to produce

In one embodiment, provided herein are methods of preparing a compoundhaving the formula:

the methods comprising contacting

with a base and ROD/D₂O, wherein said contacting occurs under conditionssuitable to produce

5.4 Pharmaceutical Compositions

In one embodiment, provided herein are pharmaceutical compositionscomprising Compound A and one or more pharmaceutically acceptableexcipients or carriers. In one embodiment, the pharmaceuticalcompositions provided herein comprise Form A of Compound A and one ormore pharmaceutically acceptable excipients or carriers. In oneembodiment, the pharmaceutical compositions provided herein compriseForm B (a hydrate) of Compound A and one or more pharmaceuticallyacceptable excipients or carriers. In one embodiment, the pharmaceuticalcompositions provided herein comprise Form C (anhydrous) of Compound Aand one or more pharmaceutically acceptable excipients or carriers. Inone embodiment, the pharmaceutical compositions provided herein compriseForm D (a methanol solvate) of Compound A and one or morepharmaceutically acceptable excipients or carriers. In one embodiment,the pharmaceutical compositions provided herein comprise Form E (ap-xylene solvate) of Compound A and one or more pharmaceuticallyacceptable excipients or carriers. In one embodiment, the pharmaceuticalcompositions provided herein comprise amorphous Compound A and one ormore pharmaceutically acceptable excipients or carriers.

In one embodiment, the pharmaceutical compositions provided hereincomprise an isotopologue of Compound A and one or more pharmaceuticallyacceptable excipients or carriers. In one embodiment, the pharmaceuticalcompositions provided herein comprise a metabolite of Compound A and oneor more pharmaceutically acceptable excipients or carriers. With respectto the pharmaceutical compositions provided herein, each reference to“Compound A” is contemplated as including pharmaceutically acceptablesalts, solid forms (including Form A, Form B, Form C, Form D, Form E,and/or amorphous Compound A), isotopologues and metabolites of CompoundA.

In one embodiment, the pharmaceutically acceptable excipients andcarriers are selected from binders, diluents, disintegrants andlubricants.

In certain embodiments, the binders include, but are not limited to,cellulose (e.g., microcrystalline cellulose, such as AVICEL® PH 101,AVICEL® PH 102 and AVICEL® PH 112) and starch (e.g., pregelatinizedstarch (STARCH 1500®)). In one embodiment, the binder is cellulose. Inanother embodiment, the binder is microcrystalline cellulose. In yetanother embodiment, the binder is AVICEL® PH 101. In yet anotherembodiment, the binder is AVICEL® PH 102. In yet another embodiment, thebinder is AVICEL® PH 112. In yet another embodiment, the binder isstarch. In yet another embodiment, the binder is pregelatinized starch.In still another embodiment, the binder is STARCH 1500®.

In certain embodiments, the diluents include, but are not limited to,lactose (e.g., lactose monohydrate (FAST FLO® 316) and lactoseanhydrous), cellulose (e.g., microcrystalline cellulose, such as AVICEL®PH 101, AVICEL® PH 102 and AVICEL® PH 112). In one embodiment, thediluent is lactose. In another embodiment, the diluent is lactosemonohydrate. In yet another embodiment, the diluent is FAST FLO® 316. Inyet another embodiment, the diluent is lactose anhydrous. In yet anotherembodiment, the diluent is cellulose. In yet another embodiment, thediluent is microcrystalline cellulose. In yet another embodiment, thediluent is AVICEL® PH 101. In still another embodiment, the diluent isAVICEL® PH 102. In still another embodiment, the diluent is AVICEL® PH112.

In certain embodiments, the disintegrants include, but are not limitedto, starch (e.g., corn starch) and carboxymethyl cellulose (e.g.,croscarmellose sodium, such as AC-DI-SOL®). In one embodiment, thedisintegrant is starch. In another embodiment, the disintegrant is cornstarch. In yet another embodiment, the disintegrant is carboxymethylcellulose. In yet another embodiment, the disintegrant is croscarmellosesodium. In still another embodiment, the disintegrant is AC-DI-SOL®.

In certain embodiments, the lubricants include, but are not limited to,starch (e.g., corn starch), magnesium stearate, and stearic acid. In oneembodiment, the lubricant is starch. In another embodiment, thelubricant is corn starch. In yet another embodiment, the lubricant ismagnesium stearate. In still another embodiment, the lubricant isstearic acid.

In another embodiment, the pharmaceutical compositions provided hereincomprise Compound A and one or more pharmaceutically acceptableexcipients or carriers, each independently selected from carboxymethylcellulose, cellulose, lactose, magnesium stearate, starch, and stearicacid.

In another embodiment, the pharmaceutical compositions provided hereincomprise Compound A and one or more pharmaceutically acceptableexcipients or carriers, each independently selected from carboxymethylcellulose, cellulose, lactose, magnesium stearate and starch.

In yet another embodiment, the pharmaceutical compositions providedherein comprise Compound A and one or more pharmaceutically acceptableexcipients or carriers, each independently selected from croscarmellosesodium, microcrystalline cellolose, lactose anhydrous, lactosemonohydrate, magnesium stearate, corn starch, pregelatinized starch, andstearic acid.

In yet another embodiment, the pharmaceutical compositions providedherein comprise Compound A and one or more pharmaceutically acceptableexcipients or carriers, each independently selected from croscarmellosesodium, microcrystalline cellolose, lactose anhydrous, lactosemonohydrate, magnesium stearate, corn starch and pregelatinized starch.

In certain embodiments, the pharmaceutical compositions provided hereindo not comprise stearic acid.

In yet another embodiment, the pharmaceutical compositions providedherein comprise Compound A and one or more pharmaceutically acceptableexcipients or carriers, each independently selected from AC-DI-SOL®,AVICEL PH 101®, AVICEL PH 102®, lactose anhydrous, FAST FLO 316®,magnesium stearate, corn starch, STARCH 1500®, and stearic acid.

In yet another embodiment, the pharmaceutical compositions providedherein comprise Compound A and one or more pharmaceutically acceptableexcipients or carriers, each independently selected from AC-DI-SOL®,AVICEL PH 101®, AVICEL PH 102®, lactose anhydrous, FAST FLO 316®,magnesium stearate, corn starch and STARCH 1500®.

In one embodiment, the pharmaceutical compositions provided hereincomprise Compound A, a diluent(s)/binder(s), a disintegrant(s), and alubricant(s).

In one embodiment, the pharmaceutical compositions provided hereincomprise Compound A, stearic acid and lactose monohydrate.

In one embodiment, the pharmaceutical compositions provided hereincomprise Compound A and lactose monohydrate.

In one embodiment, the pharmaceutical compositions provided hereincomprise Compound A, stearic acid, lactose monohydrate andmicrocyrstalline cellulose.

In one embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose monohydrate and microcyrstalline cellulose.

In another embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose monohydrate, microcrystalline cellulose,carboxymethyl cellulose, and magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose monohydrate, microcrystalline cellulose,croscarmellose sodium, stearic acid and magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose monohydrate, microcrystalline cellulose,croscarmellose sodium and magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise Compound A, FAST FLO 316®, AVICEL PH 102®, AC-DI-SOL®,stearic acid and magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise Compound A, FAST FLO 316®, AVICEL PH 112®, AC-DI-SOL®and magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise about 10-20% by weight of Compound A, about 70-90% by weight ofdiluent(s)/binder(s), about 1-5% by weight of disintegrant(s), and about0.1-2% by weight of lubricant(s).

In one embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Compound A, about 80% by weight ofdiluent(s)/binder(s), about 3% by weight of disintegrant(s), and about1.4% by weight of lubricant(s).

In another embodiment, the pharmaceutical compositions provided hereincomprise about 10-20% by weight of Form A of Compound A, about 30-60% byweight of lactose, about 20-40% by weight of microcrystalline cellulose,about 1-5% by weight of carboxymethyl cellulose, about 0.1-2% by weightof stearic acid and about 0.5-3% by weight of magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 10-20% by weight of Form A of Compound A, about 30-60% byweight of lactose, about 20-40% by weight of microcrystalline cellulose,about 1-5% by weight of carboxymethyl cellulose and about 0.5-3% byweight of magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Form A of Compound A, about 49% byweight of lactose, about 31% by weight of microcrystalline cellulose,about 3% by weight of carboxymethyl cellulose, about 0.4% by weight ofstearic acid and about 1% by weight of magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Form A of Compound A, about 49% byweight of lactose, about 31% by weight of microcrystalline cellulose,about 3% by weight of carboxymethyl cellulose and about 1% by weight ofmagnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 10-20% by weight of Form A of Compound A, about30-60% by weight of lactose monohydrate, about 20-40% by weight ofmicrocrystalline cellulose, about 1-5% by weight of croscarmellosesodium, about 0.1-2% by weight stearic acid and about 0.5-3% by weightof magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 10-20% by weight of Form A of Compound A, about30-60% by weight of lactose monohydrate, about 20-40% by weight ofmicrocrystalline cellulose, about 1-5% by weight of croscarmellosesodium and about 0.5-3% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Form A of Compound A, about 49%by weight of lactose monohydrate, about 31% by weight ofmicrocrystalline cellulose, about 3% by weight of croscarmellose sodium,about 0.4% by weight of stearic acid and about 1% by weight of magnesiumstearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Form A of Compound A, about 49%by weight of lactose monohydrate, about 31% by weight ofmicrocrystalline cellulose, about 3% by weight of croscarmellose sodiumand about 1% by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 10-20% by weight of Form A of Compound A, about30-60% by weight of FAST FLO 316®, about 20-40% by weight of AVICEL PH102®, about 1-5% by weight of AC-DI-SOL®, about 0.1-2% by weight ofstearic acid and about 0.5-3% by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 10-20% by weight of Form A of Compound A, about30-60% by weight of FAST FLO 316®, about 20-40% by weight of AVICEL PH112®, about 1-5% by weight of AC-DI-SOL® and about 0.5-3% by weight ofmagnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Form A of Compound A, about 49%by weight of FAST FLO 316®, about 31% by weight of AVICEL PH 102®, about3% by weight of AC-DI-SOL®, about 0.4% by weight of stearic acid andabout 1% by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Form A of Compound A, about 49%by weight of FAST FLO 316®, about 31% by weight of AVICEL PH 112®, about3% by weight of AC-DI-SOL® and about 1% by weight of magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise Form A of Compound A, lactose, starch, carboxymethyl cellulose,stearic acid and magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise Form A of Compound A, lactose, starch, carboxymethyl celluloseand magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise Form A of Compound A, lactose monohydrate, pregelatinizedstarch, croscarmellose sodium, stearic acid and magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise Form A of Compound A, lactose monohydrate, pregelatinizedstarch, croscarmellose sodium and magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise Compound A, FAST FLO 316®, STARCH 1500®, AC-DI-SOL®,stearic acid and magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise Compound A, FAST FLO 316®, STARCH 1500®, AC-DI-SOL® andmagnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Compound A, from about 55% to about 80%by weight of diluent(s)/binder(s), from about 20% to about 30% by weightof disintegrant(s), and about 1% by weight of lubricant(s).

In another embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Compound A, about 55% by weight oflactose, about 25% by weight of starch, about 3% by weight ofcarboxymethyl cellulose, about 0.4% by weight of stearic acid and about1% by weight of magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Compound A, about 55% by weight oflactose, about 25% by weight of starch, about 3% by weight ofcarboxymethyl cellulose and about 1% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 55% by weightof lactose monohydrate, about 25% by weight of pregelatinized starch,about 3% by weight of croscarmellose sodium, about 0.4% by weight ofstearic acid and about 1% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 55% by weightof lactose monohydrate, about 25% by weight of pregelatinized starch,about 3% by weight of croscarmellose sodium and about 1% by weight ofmagnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 55% by weightof FAST FLO 316®, about 25% by weight of STARCH 1500®, about 3% byweight of AC-DI-SOL®, about 0.4% by weight of stearic acid and about 1%by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 55% by weightof FAST FLO 316®, about 25% by weight of STARCH 1500®, about 3% byweight of AC-DI-SOL® and about 1% by weight of magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose, microcrystalline cellulose, carboxymethylcellulose, stearic acid and magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose, microcrystalline cellulose, carboxymethylcellulose and magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose monohydrate, microcrystalline cellulose,croscarmellose sodium, stearic acid and magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose monohydrate, microcrystalline cellulose,croscarmellose sodium and magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise Compound A, FAST FLO 316®, AVICEL PH 102®, AC-DI-SOL®,about 0.4% by weight of stearic acid and magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise Compound A, FAST FLO 316®, AVICEL PH 112®, AC-DI-SOL®and magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Compound A, about 80% by weight ofdiluent(s)/binder(s), about 3% by weight of disintegrant(s), and about1% by weight of lubricant(s).

In another embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Compound A, about 50% by weight oflactose, about 30% by weight of microcrystalline cellulose, about 3% byweight of carboxymethyl cellulose, about 0.4% by weight of stearic acidand about 1% by weight of magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Compound A, about 50% by weight oflactose, about 30% by weight of microcrystalline cellulose, about 3% byweight of carboxymethyl cellulose and about 1% by weight of magnesiumstearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 50% by weightof lactose monohydrate, about 30% by weight of microcrystallinecellulose, about 3% by weight of croscarmellose sodium, about 0.4% byweight of stearic acid and about 1% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 50% by weightof lactose monohydrate, about 30% by weight of microcrystallinecellulose, about 3% by weight of croscarmellose sodium and about 1% byweight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 50% by weightof FAST FLO 316®, about 30% by weight of AVICEL PH 102®, about 3% byweight of AC-DI-SOL®, about 0.4% by weight of stearic acid and about 1%by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 50% by weightof FAST FLO 316®, about 30% by weight of AVICEL PH 112®, about 3% byweight of AC-DI-SOL® and about 1% by weight of magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose, microcrystalline cellulose, corn starch,carboxymethyl cellulose, stearic acid and magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose, microcrystalline cellulose, corn starch,carboxymethyl cellulose and magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose monohydrate, microcrystalline cellulose,corn starch, croscarmellose sodium, stearic acid and magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose monohydrate, microcrystalline cellulose,corn starch, croscarmellose sodium and magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise Compound A, FAST FLO 316®, AVICEL PH 102®, corn starch,AC-DI-SOL®, stearic acid and magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise Compound A, FAST FLO 316®, AVICEL PH 102®, corn starch,AC-DI-SOL® and magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Compound A, from about 85% to about 90%by weight of diluent(s)/binder(s), from about 1% to about 10% by weightof disintegrant(s), and from about 1% to about 6% by weight oflubricants.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Compound A, about 45% by weight oflactose, about 30% by weight of microcrystalline cellulose, about 3% byweight of corn starch, about 3% by weight of carboxymethyl cellulose,about 0.4% by weight of stearic acid and about 1% by weight of magnesiumstearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Compound A, about 45% by weight oflactose, about 30% by weight of microcrystalline cellulose, about 3% byweight of corn starch, about 3% by weight of carboxymethyl cellulose andabout 1% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 88% by weightof lactose, about 25% by weight of microcrystalline cellulose, about 4%by weight of corn starch, about 4% by weight of carboxymethyl cellulose,about 0.4% by weight of stearic acid and about 1.5% by weight ofmagnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 88% by weightof lactose, about 25% by weight of microcrystalline cellulose, about 4%by weight of corn starch, about 4% by weight of carboxymethyl celluloseand about 1.5% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 45% by weightof lactose monohydrate, about 30% by weight of microcrystallinecellulose, about 3% by weight of corn starch, about 3% by weight ofcroscarmellose sodium, about 0.4% by weight of stearic acid and about 1%by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 45% by weightof lactose monohydrate, about 30% by weight of microcrystallinecellulose, about 3% by weight of corn starch, about 3% by weight ofcroscarmellose sodium and about 1% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 88% by weightof lactose monohydrate, about 25% by weight of microcrystallinecellulose, about 4% by weight of corn starch, about 4% by weight ofcroscarmellose sodium, about 0.4% by weight of stearic acid and about1.5% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 88% by weightof lactose monohydrate, about 25% by weight of microcrystallinecellulose, about 4% by weight of corn starch, about 4% by weight ofcroscarmellose sodium and about 1.5% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 45% by weightof FAST FLO 316®, about 30% by weight of AVICEL PH 102®, about 3% byweight of corn starch, about 3% by weight of AC-DI-SOL®, about 0.4% byweight of stearic acid and about 1% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 45% by weightof FAST FLO 316®, about 30% by weight of AVICEL PH 102®, about 3% byweight of corn starch, about 3% by weight of AC-DI-SOL® and about 1% byweight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 88% by weightof FAST FLO 316®, about 25% by weight of AVICEL PH 102®, about 4% byweight of corn starch, about 4% by weight of AC-DI-SOL®, about 0.4% byweight of stearic acid and about 1.5% by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 88% by weightof FAST FLO 316®, about 25% by weight of AVICEL PH 102®, about 4% byweight of corn starch, about 4% by weight of AC-DI-SOL® and about 1.5%by weight of magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose, microcrystalline cellulose, corn starch,carboxymethyl cellulose, stearic acid, and magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose, microcrystalline cellulose, corn starch,carboxymethyl cellulose and magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise about 5% by weight of Compound A, about 90% by weight ofdiluent(s)/binder(s), from about 3% to about 6% by weight ofdisintegrant(s), and from about 1.5% to about 5% by weight oflubricants.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 5% by weight of Compound A, about 60% by weight oflactose, about 30% by weight of microcrystalline cellulose, about 3% byweight of corn starch, about 3% by weight of carboxymethyl cellulose,about 0.5% by weight of stearic acid, and about 1% by weight ofmagnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 5% by weight of Compound A, about 60% by weight oflactose, about 30% by weight of microcrystalline cellulose, about 3% byweight of corn starch, about 3% by weight of carboxymethyl cellulose andabout 1% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 5% by weight of Compound A, about 60% by weight oflactose monohydrate, about 30% by weight of microcrystalline cellulose,about 3% by weight of corn starch, about 3% by weight of croscarmellosesodium, about 0.5% by weight of stearic acid, and about 1% by weight ofmagnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 5% by weight of Compound A, about 60% by weight oflactose monohydrate, about 30% by weight of microcrystalline cellulose,about 3% by weight of corn starch, about 3% by weight of croscarmellosesodium and about 1% by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 5% by weight of Compound A, about 60% by weight ofFAST FLO 316®, about 30% by weight of AVICEL PH 102®, about 3% by weightof corn starch, about 3% by weight of AC-DI-SOL®, about 0.5% by weightof stearic acid, and about 1% by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 5% by weight of Compound A, about 60% by weight ofFAST FLO 316®, about 30% by weight of AVICEL PH 102®, about 3% by weightof corn starch, about 3% by weight of AC-DI-SOL® and about 1% by weightof magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose, microcrystalline cellulose, carboxymethylcellulose, stearic acid, and magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose, microcrystalline cellulose, carboxymethylcellulose and magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose monohydrate, microcrystalline cellulose,croscarmellose sodium, stearic acid, and magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise Compound A, lactose monohydrate, microcrystalline cellulose,croscarmellose sodium and magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise Compound A, FAST FLO 316®, AVICEL PH 102®, AC-DI-SOL®,stearic acid, and magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise Compound A, FAST FLO 316®, AVICEL PH 102®, AC-DI-SOL®and magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise about 12% by weight of Compound A, from about 80% to about 85%by weight of diluent(s)/binder(s), about 3% by weight ofdisintegrant(s), and about 1.5% by weight of lubricant(s).

In another embodiment, the pharmaceutical compositions provided hereincomprise about 12% by weight of Compound A, about 52.5% by weight oflactose, about 30% by weight of microcrystalline cellulose, about 3% byweight of carboxymethyl cellulose, about 0.5% by weight of stearic acid,and about 1% by weight of magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 12% by weight of Compound A, about 52.5% by weight oflactose, about 30% by weight of microcrystalline cellulose, about 3% byweight of carboxymethyl cellulose and about 1% by weight of magnesiumstearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 12% by weight of Compound A, about 52.5% by weightof lactose monohydrate, about 30% by weight of microcrystallinecellulose, about 3% by weight of croscarmellose sodium, about 0.5% byweight of stearic acid, and about 1% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 12% by weight of Compound A, about 52.5% by weightof lactose monohydrate, about 30% by weight of microcrystallinecellulose, about 3% by weight of croscarmellose sodium and about 1% byweight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 12% by weight of Compound A, about 52.5% by weightof FAST FLO 316®, about 30% by weight of AVICEL PH 102®, about 3% byweight of AC-DI-SOL®, about 0.5% by weight of stearic acid, and about 1%by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 12% by weight of Compound A, about 52.5% by weightof FAST FLO 316®, about 30% by weight of AVICEL PH 102®, about 3% byweight of AC-DI-SOL® and about 1% by weight of magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise about 12% by weight of Compound A, about 80% by weight ofdiluent(s)/binder(s), about 3% by weight of disintegrant(s), and about4% by weight of lubricant(s).

In another embodiment, the pharmaceutical compositions provided hereincomprise about 12% by weight of Compound A, about 63% by weight oflactose, about 18% by weight of microcrystalline cellulose, about 3% byweight of carboxymethyl cellulose, about 3% by weight of stearic acid,and about 1% by weight of magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 12% by weight of Compound A, about 63% by weight oflactose, about 18% by weight of microcrystalline cellulose, about 3% byweight of carboxymethyl cellulose and about 1% by weight of magnesiumstearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 12% by weight of Compound A, about 63% by weightof lactose monohydrate, about 18% by weight of microcrystallinecellulose, about 3% by weight of croscarmellose sodium, about 3% byweight of stearic acid, and about 1% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 12% by weight of Compound A, about 63% by weightof lactose monohydrate, about 18% by weight of microcrystallinecellulose, about 3% by weight of croscarmellose sodium and about 1% byweight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 12% by weight of Compound A, about 63% by weightof FAST FLO 316®, about 18% by weight of AVICEL PH 102®, about 3% byweight of AC-DI-SOL®, about 3% by weight of stearic acid, and about 1%by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 12% by weight of Compound A, about 63% by weightof FAST FLO 316®, about 18% by weight of AVICEL PH 102®, about 3% byweight of AC-DI-SOL® and about 1% by weight of magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Compound A, about 80% by weight of adiluent/binder, about 3% by weight of a disintegrant, and about 1.5% byweight of lubricants.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Compound A, about 50% by weight oflactose, about 30% by weight of microcrystalline cellulose, about 3% byweight of carboxymethyl cellulose, about 0.5% by weight of stearic acid,and about 1% by weight of magnesium stearate.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 15% by weight of Compound A, about 50% by weight oflactose, about 30% by weight of microcrystalline cellulose, about 3% byweight of carboxymethyl cellulose and about 1% by weight of magnesiumstearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 50% by weightof lactose monohydrate, about 30% by weight of microcrystallinecellulose, about 3% by weight of croscarmellose sodium, about 0.5% byweight of stearic acid, and about 1% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 50% by weightof lactose monohydrate, about 30% by weight of microcrystallinecellulose, about 3% by weight of croscarmellose sodium and about 1% byweight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 50% by weightof FAST FLO 316®, about 30% by weight of AVICEL PH 102®, about 3% byweight of AC-DI-SOL®, about 0.5% by weight of stearic acid, and about 1%by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 15% by weight of Compound A, about 50% by weightof FAST FLO 316®, about 30% by weight of AVICEL PH 102®, about 3% byweight of AC-DI-SOL® and about 1% by weight of magnesium stearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise about 17% by weight of Form A of Compound A, about 80% byweight of diluent(s)/binder(s), about 3% by weight of disintegrant(s),and about 1% by weight of lubricant(s).

In another embodiment, the pharmaceutical compositions provided hereincomprise about 17% by weight of Form A of Compound A, about 50% byweight of lactose, about 30% by weight of microcrystalline cellulose,about 3% by weight of carboxymethyl cellulose, and about 1% by weight ofmagnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 17% by weight of Form A of Compound A, about 50%by weight of lactose monohydrate, about 30% by weight ofmicrocrystalline cellulose, about 3% by weight of croscarmellose sodium,and about 1% by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 17% by weight of Form A of Compound A, about 50%by weight of FAST FLO 316®, about 30% by weight of AVICEL PH 101®, about3% by weight of AC-DI-SOL®, and about 1% by weight of magnesiumstearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise about 17% by weight of Form A of Compound A, from about 55% toabout 80% by weight of diluent(s)/binder(s), from about 20% to about 30%by weight of disintegrant(s), and about 1% by weight of lubricant(s).

In another embodiment, the pharmaceutical compositions provided hereincomprise about 17% by weight of Form A of Compound A, about 55% byweight of lactose, about 25% by weight of starch, about 3% by weight ofcarboxymethyl cellulose, and about 1% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 17% by weight of Form A of Compound A, about 55%by weight of lactose monohydrate, about 25% by weight of pregelatinizedstarch, about 3% by weight of croscarmellose sodium, and about 1% byweight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 17% by weight of Form A of Compound A, about 55%by weight of FAST FLO 316®, about 25% by weight of STARCH 1500®, about3% by weight of AC-DI-SOL®, and about 1% by weight of magnesiumstearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise about 17% by weight of Form A of Compound A, about 80% byweight of diluent(s)/binder(s), about 3% by weight of disintegrant(s),and about 1% by weight of lubricant(s).

In another embodiment, the pharmaceutical compositions provided hereincomprise about 17% by weight of Form A of Compound A, about 50% byweight of lactose, about 30% by weight of microcrystalline cellulose,about 3% by weight of carboxymethyl cellulose, and about 1% by weight ofmagnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 17% by weight of Form A of Compound A, about 50%by weight of lactose monohydrate, about 30% by weight ofmicrocrystalline cellulose, about 3% by weight of croscarmellose sodium,and about 1% by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 17% by weight of Form A of Compound A, about 50%by weight of FAST FLO 316®, about 30% by weight of AVICEL PH 102®, about3% by weight of AC-DI-SOL®, and about 1% by weight of magnesiumstearate.

In one embodiment, the pharmaceutical compositions provided hereincomprise about 17% by weight of Form A of Compound A, from about 85% toabout 90% by weight of diluent(s)/binder(s), from about 3% to about 9%by weight of disintegrant(s), and from about 1% to about 6% by weight oflubricants.

In another embodiment, the pharmaceutical compositions provided hereincomprise about 17% by weight of Form A of Compound A, about 45% byweight of lactose, about 30% by weight of microcrystalline cellulose,about 3% by weight of corn starch, about 3% by weight of carboxymethylcellulose, and about 1% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 17% by weight of Form A of Compound A, about 88%by weight of lactose, about 25% by weight of microcrystalline cellulose,about 4% by weight of corn starch, about 4% by weight of carboxymethylcellulose, and about 1.5% by weight of magnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 17% by weight of Form A of Compound A, about 45%by weight of lactose monohydrate, about 30% by weight ofmicrocrystalline cellulose, about 3% by weight of corn starch, about 3%by weight of croscarmellose sodium, and about 1% by weight of magnesiumstearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 17% by weight of Form A of Compound A, about 88%by weight of lactose monohydrate, about 25% by weight ofmicrocrystalline cellulose, about 4% by weight of corn starch, about 4%by weight of croscarmellose sodium, and about 1.5% by weight ofmagnesium stearate.

In yet another embodiment, the pharmaceutical compositions providedherein comprise about 17% by weight of Form A of Compound A, about 45%by weight of FAST FLO 316®, about 30% by weight of AVICEL PH 102®, about3% by weight of corn starch, about 3% by weight of AC-DI-SOL®, and about1% by weight of magnesium stearate.

In still another embodiment, the pharmaceutical compositions providedherein comprise about 17% by weight of Form A of Compound A, about 88%by weight of FAST FLO 316®, about 25% by weight of AVICEL PH 102®, about4% by weight of corn starch, about 4% by weight of AC-DI-SOL®, and about1.5% by weight of magnesium stearate.

In certain embodiments, provided herein are pharmaceutical compositionscomprising Compound A and stearic acid. In certain embodiments, stearicacid is present in an amount of about 0.1-5%, 0.1 to 1%, or 0.4% byweight. Without being limited by theory, it was found that the additionof stearic acid improved lubrication (reduced sticking) withoutimpacting disintegration and compressibility.

In certain embodiments, provided herein are pharmaceutical compositionscomprising Compound A and lactose monohydrate. In certain embodiments,lactose monohydrate is present in an amount of about 40-60%, 45-55%,49.2% or 49.6% by weight. Without being limited by theory, it was foundthat lactose monohydrate provided better flowability than lactoseanhydrous.

In certain embodiments, provided herein are pharmaceutical compositionscomprising Compound A and AVICEL PH 102®. In certain embodiments, AVICELPH 102® is present in an amount of about 20-40%, 25-35%, or 31% byweight. Without being limited by theory, it was found that AVICEL PH102® provided better flowability than AVICEL PH 101®.

In certain embodiments, provided herein are pharmaceutical compositionscomprising Compound A and AVICEL PH 112®. In certain embodiments, AVICELPH 112® is present in an amount of about 20-40%, about 25-35%, or about31% by weight. It was unexpectedly found that Compound A is susceptibleto hydrolysis. Without being limited by theory, it is thought thatAVICEL PH 112®, being a low-moisture grade microcrystalline cellulose,can reduce hydrolysis of Compound A.

In certain embodiments, provided herein are pharmaceutical compositionscomprising Compound A, stearic acid, lactose monohydrate and AVICEL PH102®. In certain embodiments, provided herein are pharmaceuticalcompositions comprising Compound A, stearic acid (in an amount of about0.1-5%, 0.1 to 1%, or 0.4% by weight), lactose monohydrate (in an amountof about 40-60%, 45-55%, or 49.2% by weight) and AVICEL PH 102® (in anamount of about 20-40%, 25-35%, or 31% by weight).

In certain embodiments, provided herein are pharmaceutical compositionscomprising Compound A, lactose monohydrate and AVICEL PH 102®. Incertain embodiments, provided herein are pharmaceutical compositionscomprising Compound A, lactose monohydrate (in an amount of about40-60%, 45-55%, or 49.2% by weight) and AVICEL PH 102® (in an amount ofabout 20-40%, 25-35%, or 31% by weight).

In certain embodiments, provided herein are pharmaceutical compositionscomprising an opaque coating. Without being limited by theory, it wasfound that a more opaque coating protected the drug product fromdegradation. In some embodiments, the pharmaceutical composition isformulated as a tablet. In some such embodiments, the tablet is filmcoated. In some embodiments, the tablet is film coated to a weight gainof 1-8%. In others, the film coating is about 4% by weight of thetablet.

In certain embodiments, provided herein are pharmaceutical compositionscomprising Compound A that do not comprise stearic acid. Without beinglimited by theory, a lack of picking or sticking of certain tabletformulations by visual observation indicated that acceptable tabletformulations could be produced without the use of stearic acid.

In certain embodiments, provided herein are pharmaceutical compositionsas set forth in Table 6-Table 14, Table 17-Table 19, Table 26-Table 28and Table 31-Table 34, wherein the amounts of the recited components canindependently be varied by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%,20% or 25%.

In certain embodiments, provided herein are liquid formulationscomprising Compound A, an alcohol and polyethylene glycol. In certainembodiments, the alcohol and polyethylene glycol are present in a ratioof about 80:20 to about 20:80. In certain embodiments, the alcohol andpolyethylene glycol are present in a ratio of about 50:50. In certainembodiments, the alcohol is ethanol. In certain embodiments, thepolyethylene glycol is PEG 400. In one embodiment, provided herein arecapsules filled with a liquid formulation comprising Compound A, analcohol and polyethylene glycol. In one embodiment, Compound A is anisotopologue of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.In some embodiments, the isotopologue is enriched in ¹⁴C.

The pharmaceutical compositions provided herein can be provided in aunit-dosage form or multiple-dosage form. A unit-dosage form, as usedherein, refers to physically discrete unit suitable for administrationto a human and animal subject, and packaged individually as is known inthe art. Each unit-dose contains a predetermined quantity of an activeingredient(s) sufficient to produce the desired therapeutic effect, inassociation with the required pharmaceutical carriers or excipients.Examples of a unit-dosage form include an individually packaged tabletor capsule. A unit-dosage form may be administered in fractions ormultiples thereof. A multiple-dosage form is a plurality of identicalunit-dosage forms packaged in a single container to be administered insegregated unit-dosage form. In certain embodiments, the unit dosageforms provided herein comprise about 1 mg to about 100 mg of Compound A.In other embodiments, the unit dosage forms provided herein compriseabout 5 mg to about 50 mg of Compound A. In other embodiments, the unitdosage forms provided herein comprise about 1 mg, about 5 mg, about 20mg, about 45 mg, about 50 mg, about 75 mg or about 100 mg of Compound A.In other embodiments, the unit dosage forms provided herein compriseabout 5 mg, about 15 mg, about 20 mg, about 30 mg, about 45 mg, andabout 50 mg of Compound A.

In certain embodiments, provided herein are methods for preparing acomposition provided herein, comprising: (i) weighing out the desiredamount of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform (such as Form A, Form B, Form C, Form D, or Form E) thereof and thedesired amount of excipients (such as lactose monohydrate,croscarmellose sodium and microcrystalline cellulose); (ii) mixing orblending7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the excipients; (iii) passing the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and excipients through a screen (such as an 18 mesh or 1000μm screen); (iv) mixing or blending7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the excipients after passage through the screen; (v)weighing out the desired amount of lubricating agents (such as stearicacid and/or magnesium stearate); (vi) passing the lubricating agentsthrough a screen (such as a 30 mesh or 600 μm screen); (vii) mixing orblending7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents; (viii)compressing the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents (such as into atablet form); and (ix) coating the compressed mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents with a coatingagent (such as Opadry pink, yellow or beige).

In certain embodiments, provided herein are methods for preparing acomposition provided herein, comprising: (i) weighing out the desiredamount of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform (such as Form A, Form B, Form C, Form D, or Form E) thereof and thedesired amount of excipients (such as lactose monohydrate,croscarmellose sodium and microcrystalline cellulose); (ii) mixing orblending7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the excipients; (iii) passing the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and excipients through a screen (such as an 18 mesh or 1000μm screen); (iv) mixing or blending7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the excipients after passage through the screen; (v)weighing out the desired amount of lubricating agents (such as stearicacid and/or magnesium stearate); (vi) passing the lubricating agentsthrough a screen (such as a 60 mesh or 250 μm screen); (vii) mixing orblending7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents; (viii)compressing the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents (such as into atablet form); and (ix) coating the compressed mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents with a coatingagent (such as Opadry pink, yellow or beige).

In certain embodiments, provided herein are methods for preparing acomposition provided herein, comprising: (i) weighing out the desiredamount of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the desired amount of excipients (such as lactosemonohydrate, croscarmellose sodium and microcrystalline cellulose); (ii)passing the excipients through a screen (such as an 18 mesh or 1000 μmscreen); (iii) mixing or blending (such as at 26 revolutions per minutefor 20 minutes)7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform (such as Form A, Form B, Form C, Form D, or Form E) thereof and theexcipients; (iv) passing the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and excipients through a screen (such as an 18 mesh or 1000μm screen); (v) mixing or blending (such as at 26 revolutions per minutefor 10 minutes)7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the excipients; (vi) weighing out the desired amount oflubricating agents (such as stearic acid and/or magnesium stearate);(vii) passing the lubricating agents through a screen (such as a 30 meshor 600 μm screen); (viii) mixing or blending (such as at 26 revolutionsper minute for 3 minutes)7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents; (ix)compressing the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents (such as into atablet form); and (x) coating the compressed mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents with a coatingagent (such as Opadry pink, yellow or beige).

In certain embodiments, provided herein are methods for preparing acomposition provided herein, comprising: (i) weighing out the desiredamount of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydro-pyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the desired amount of excipients (such as lactosemonohydrate, croscarmellose sodium and microcrystalline cellulose); (ii)passing the excipients through a screen (such as an 18 mesh or 1000 μmscreen); (iii) mixing or blending (such as at 26 revolutions per minutefor 20 minutes)7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform (such as Form A, Form B, Form C, Form D, or Form E) thereof and theexcipients; (iv) passing the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and excipients through a screen (such as an 18 mesh or 1000μm screen); (v) mixing or blending (such as at 26 revolutions per minutefor 10 minutes)7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof and the excipients; (vi) weighing out the desired amount oflubricating agents (such as stearic acid and/or magnesium stearate);(vii) passing the lubricating agents through a screen (such as a 60 meshor 250 μm screen); (viii) mixing or blending (such as at 26 revolutionsper minute for 3 minutes)7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents; (ix)compressing the mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents (such as into atablet form); and (x) coating the compressed mixture of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a pharmaceutically acceptable salt, isotopologue, metabolite or solidform thereof, the excipients and the lubricating agents with a coatingagent (such as Opadry pink, yellow or beige).

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form A of Compound A, including substantially pure Form A.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form B of Compound A, including substantially pure Form B.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form C of Compound A, including substantially pure Form C.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form D of Compound A, including substantially pure Form D.

In certain embodiments, the pharmaceutical compositions provided hereincomprise Form E of Compound A, including substantially pure Form E.

Further provided herein are kits comprising a pharmaceutical compositionof Compound A provided herein. In particular embodiments, providedherein are kits comprising a unit dosage form of Compound A providedherein. In certain embodiments of the kits provided herein, Compound Ais provided as Form A. In certain embodiments of the kits providedherein, Compound A is provided as Form B. In certain embodiments of thekits provided herein, Compound A is provided as Form C. In certainembodiments of the kits provided herein, Compound A is provided as FormD. In certain embodiments of the kits provided herein, Compound A isprovided as Form E. In certain embodiments of the kits provided herein,Compound A is provided as a pinacol co-crystal. In certain embodimentsof the kits provided herein, Compound A is provided as an amorphousform. In some embodiments, of the kits provided herein Compound A isprovided as an isotopologue of7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one.In some such embodiments, the isotopologue is enriched in is enriched in¹³C, ¹⁴C, ²H, ³H and/or ¹⁵N.

5.5 Methods of Use

The solid forms of Compound A (e.g., Form A, Form B, Form C, Form D,Form E or amorphous), isotopologues of Compound A, metabolites ofCompound A (e.g, O-desmethyl Compound A) and the pharmaceuticalcompositions provided herein have utility as pharmaceuticals to treat orprevent a disease in a subject, e.g., a proliferative disease. Further,the solid forms of Compound A (e.g., Form A, Form B, Form C, Form D,Form E or amorphous), isotopologues of Compound A, metabolites ofCompound A (e.g, O-desmethyl Compound A) and the pharmaceuticalcompositions provided herein provided herein are active against kinases(e.g., protein kinases), including those involved in cancer,inflammatory conditions, immunological conditions, neurodegenerativediseases, diabetes, obesity, neurological disorders, age-relateddiseases, and/or cardiovascular conditions. Without being limited bytheory, it is thought the solid forms of Compound A (e.g., Form A, FormB, Form C, Form D, Form E or amorphous), isotopologues of Compound A,metabolites of Compound A (e.g, O-desmethyl Compound A) and thepharmaceutical compositions provided herein are effective for treatingand preventing diseases and conditions due to its ability to modulate(e.g., inhibit) kinases that are involved in the etiology of thediseases and conditions. Accordingly, provided herein are uses of thesolid forms of Compound A (e.g., Form A, Form B, Form C, Form D, Form Eor amorphous), isotopologues of Compound A, metabolites of Compound A(e.g, O-desmethyl Compound A) and the pharmaceutical compositionsprovided herein, including the treatment or prevention of those diseasesset forth herein. In certain embodiments, the methods provided hereincomprise administering a solid form of Compound A (e.g., Form A, Form B,Form C, Form D, Form E or amorphous), an isotopologue of Compound A, ametabolite of Compound A (e.g, O-desmethyl Compound A) or apharmaceutical composition provided herein, wherein the solid form ofCompound A (e.g., Form A, Form B, Form C, Form D, Form E or amorphous),isotopologue of Compound A, metabolite of Compound A (e.g, O-desmethylCompound A) or the pharmaceutical composition provided herein is part ofa kit provided herein.

In one embodiment, provided herein is a method of treating andpreventing a disease or condition in a subject, comprising theadministration of an effective amount of the solid form of Compound A(e.g., Form A, Form B, Form C, Form D, Form E or amorphous), anisotopologue of Compound A, a metabolite of Compound A (e.g, O-desmethylCompound A) or a pharmaceutical composition provided herein to thesubject.

Representative immunological conditions that the solid forms of CompoundA (e.g., Form A, Form B, Form C, Form D, Form E or amorphous),isotopologues of Compound A, metabolites of Compound A (e.g, O-desmethylCompound A) and the pharmaceutical compositions provided herein areuseful for treating or preventing include, but are not limited to,rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, multiplesclerosis, lupus, inflammatory bowel disease, ulcerative colitis,Crohn's disease, myasthenia gravis, Graves disease, encephalomyelitis,Type II diabetes, dermatomyositis, and transplant rejection (e.g., inthe treatment of recipients of heart, lung, combined heart-lung, liver,kidney, pancreatic, skin, or corneal transplants; or graft-versus-hostdisease, such as following bone marrow transplantation).

Representative inflammatory conditions that the solid forms of CompoundA (e.g., Form A, Form B, Form C, Form D, Form E or amorphous),isotopologues of Compound A, metabolites of Compound A (e.g, O-desmethylCompound A) and the pharmaceutical compositions provided herein areuseful for treating or preventing include, but are not limited to,psoriasis, asthma and allergic rhinitis, bronchitis, chronic obstructivepulmonary disease, cystic fibrosis, inflammatory bowel disease,irritable bowel syndrome, Crohn's disease, mucous colitis, ulcerativecolitis, and obesity.

Representative cardiovascular diseases that the solids form of CompoundA (e.g., Form A, Form B, Form C, Form D, Form E or amorphous),isotopologues of Compound A, metabolites of Compound A (e.g, O-desmethylCompound A) and the pharmaceutical compositions provided herein areuseful for treating or preventing include, but are not limited to,restenosis, Wolf-Parkinson-White Syndrome, stroke, myocardial infarctionor ischemic damage to the heart, lung, gut, kidney, liver, pancreas,spleen or brain.

Representative neurodegenerative diseases that the solid forms ofCompound A (e.g., Form A, Form B, Form C, Form D, Form E or amorphous),isotopologues of Compound A, metabolites of Compound A (e.g, O-desmethylCompound A) and the pharmaceutical compositions provided herein areuseful for treating or preventing include, but are not limited to,Huntington's disease, Alzheimer's disease, Parkinson's disease,dementias caused by tau mutations, spinocerebellar ataxia type 3, motorneuron disease caused by SOD1 mutations, neuronal ceroidlipofucinoses/Batten disease (pediatric neurodegene ration) andHIV-associated encephalitis.

Representative age-related diseases that the solid forms of Compound A(e.g., Form A, Form B, Form C, Form D, Form E or amorphous),isotopologues of Compound A, metabolites of Compound A (e.g, O-desmethylCompound A) and the pharmaceutical compositions provided herein areuseful for treating or preventing include, but are not limited to,cancer, obesity, type II diabetes mellitus, autoimmune disease,cardiovascular diseases and neuronal degeneration.

In certain embodiments, the disease or condition is a fibrotic diseaseor disorder. Thus, in one embodiment, provided herein is a method fortreating or preventing a fibrotic disease or disorder in a subject,comprising the administration of an effective amount of the solid formof Compound A (e.g., Form A, Form B, Form C, Form D, Form E oramorphous), an isotopologue of Compound A, a metabolite of Compound A(e.g, O-desmethyl Compound A) or a pharmaceutical composition providedherein to the subject. In another embodiment, provided herein is amethod of treating or preventing scleroderma, idiopathic pulmonaryfibrosis, renal fibrosis, cystic fibrosis, myelofibrosis, hepaticfibrosis, steatofibrosis or steatohepatitis in a subject, comprising theadministration of an effective amount of the solid form of Compound A(e.g., Form A, Form B, Form C, Form D, Form E or amorphous), anisotopologue of Compound A, a metabolite of Compound A (e.g, O-desmethylCompound A) or a pharmaceutical composition provided herein to thesubject.

Representative cancers that the solid forms of Compound A (e.g., Form A,Form B, Form C, Form D, Form E or amorphous), isotopologues of CompoundA, metabolites of Compound A (e.g, O-desmethyl Compound A) and thepharmaceutical compositions provided herein are useful for treating orpreventing include, but are not limited to, cancers of the head, neck,eye, mouth, throat, esophagus, bronchus, larynx, pharynx, chest, bone,lung, colon, rectum, stomach, prostate, urinary bladder, uterine,cervix, breast, ovaries, testicles or other reproductive organs, skin,thyroid, blood, lymph nodes, kidney, liver, pancreas, and brain orcentral nervous system. The solid forms of Compound A (e.g., Form A,Form B, Form C, Form D, Form E or amorphous), isotopologues of CompoundA, metabolites of Compound A (e.g, O-desmethyl Compound A) and thepharmaceutical compositions provided hereinare also useful for treatingor preventing solid tumors and bloodborne tumors.

In some embodiments, the cancers within the scope of the methodsprovided herein include those associated with the pathways involvingmTOR, PI3K, or Akt kinases and mutants or isoforms thereof. In someembodiments, the cancers within the scope of the methods provided hereininclude those associated with the pathways of the following kinases:PI3Kα, PI3Kβ, PI3Kδ, KDR, GSK3α, GSK3β, ATM, ATX, ATR, cFMS, and/orDNA-PK kinases and mutants or isoforms thereof. In some embodiments, thecancers associated with mTOR/PI3K/Akt pathways include solid andblood-borne tumors, for example, multiple myeloma, mantle cell lymphoma,diffused large B-cell lymphoma, acute myeloid lymphoma, follicularlymphoma, chronic lymphocytic leukemia; breast, lung, endometrial,ovarian, gastric, cervical, and prostate cancer; glioblastoma; renalcarcinoma; hepatocellular carcinoma; colon carcinoma; neuroendocrinetumors; head and neck tumors; and sarcomas.

In one embodiment, provided herein is a method for treating orpreventing a disease or disorder associated with activation of mTORsignaling, comprising the administration of an effective amount of thesolid form of Compound A (e.g., Form A, Form B, Form C, Form D, Form Eor amorphous), an isotopologue of Compound A, a metabolite of Compound A(e.g, O-desmethyl Compound A) or a pharmaceutical composition providedherein to a subject in need thereof. Examples of diseases or disordersassociated with activation of mTOR signaling include, but are notlimited to, tumor syndromes resulting directly or indirectly fromgenetic defects in PTEN (Phosphatase and tensin homologue deleted onchromosome 10), TSC1 (Tuberous sclerosis 1), TSC2 (Tuberous sclerosis2), NF1 (Neurofibromin 1), AMPK (AMP-dependent protein kinase STK11,serine/threonine kinase 11), LKB1, VHL (von Hippel-Lindau disease) andPKD1 (polycystin-1). Without being limited by theory, it is thought thatgenetic defects associated with these proteins results inhyperactivation of the mTOR/PI3K/Akt pathway. In certain embodiments,the diseases which are treatable or preventable through inhibition ofthe mTOR/PI3K/Akt pathway include, but are not limited to, Cowden'sdisease, Cowden syndrome, Cowden-like syndrome, Bannayan-Zonanasyndrome, Bannayan-Riley-Ruvalcaba syndrome, Lhermitte-Duclos disease,endometrial carcinoma, tuberous sclerosis complex,lymphangioleiomyomatosis, neurofibromatosis 1, Peutz-Jeghers syndrome,renal cell carcinoma, von Hippel-Lindau disease, Proteus syndrome, andpolycystic kidney disease.

In another embodiment, provided herein is a method for treating orpreventing a disease or disorder associated with mTOR, PI3K, Akt, and/orDNA-PK signaling, comprising the administration of an effective amountof the solid form of Compound A (e.g., Form A, Form B, Form C, Form D,Form E or amorphous), an isotopologue of Compound A, a metabolite ofCompound A (e.g, O-desmethyl Compound A) or a pharmaceutical compositionprovided hereinto a subject in need thereof. Examples of diseases whichare treatable or preventable by inhibiting mTOR, PI3K, Akt and/or DNA-PKsignaling, include, but are not limited to, rheumatoid arthritis;rheumatoid spondylitis; osteoarthritis; gout; asthma, bronchitis;allergic rhinitis; chronic obstructive pulmonary disease; cysticfibrosis; inflammatory bowel disease; irritable bowel syndrome; mucouscolitis; ulcerative colitis; Crohn's disease; Huntington's disease;gastritis; esophagitis; hepatitis; pancreatitis; nephritis; multiplesclerosis; lupus erythematosus; atherosclerosis; restenosis followingangioplasty; left ventricular hypertrophy; myocardial infarction;stroke; ischemic damages of heart, lung, gut, kidney, liver, pancreas,spleen and brain; acute or chronic organ transplant rejection;preservation of the organ for transplantation; organ failure or loss oflimb (e.g., including, but not limited to, that resulting fromischemia-reperfusion injury, trauma, gross bodily injury, car accident,crush injury or transplant failure); graft versus host disease;endotoxin shock; multiple organ failure; psoriasis; burn from exposureto fire, chemicals or radiation; eczema; dermatitis; skin graft;ischemia; ischemic conditions associated with surgery or traumaticinjury (e.g., vehicle accident, gunshot wound or limb crush); epilepsy;Alzheimer's disease; Parkinson's disease; immunological response tobacterial or viral infection; cachexia; angiogenic and proliferativediseases (including retinitis pigmentosa), solid tumors, and cancers ofa variety of tissues such as colon, rectum, prostate, liver, lung,bronchus, pancreas, brain, head, neck, stomach, skin, kidney, cervix,blood, larynx, esophagus, mouth, pharynx, urinary bladder, ovary oruterine.

In yet another embodiment, provided herein is a method of inhibiting akinase in a cell expressing the kinase, comprising contacting the cellwith an effective amount of the solid form of Compound A (e.g., Form A,Form B, Form C, Form D, Form E or amorphous), an isotopologue ofCompound A, a metabolite of Compound A (e.g, O-desmethyl Compound A) ora pharmaceutical composition provided herein provided herein. In oneembodiment, the kinase is TOR kinase. In certain embodiments, the cellis in a subject. In certain embodiments, the cell is from a subject.

In yet another embodiment, provided herein is a method of treating orpreventing a condition treatable or preventable by the inhibition of akinase pathway, in one embodiment, the mTOR/PI3K/Akt and/or DNA-PKpathway, comprising administering to a subject in need thereof aneffective amount of the solid form of Compound A (e.g., Form A, Form B,Form C, Form D, Form E or amorphous), an isotopologue of Compound A, ametabolite of Compound A (e.g, O-desmethyl Compound A) or apharmaceutical composition provided herein. Conditions treatable orpreventable by the inhibition of the mTOR/PI3K/Akt pathway include, butare not limited to, solid and blood-borne tumors, for example, multiplemyeloma, mantle cell lymphoma, diffused large B-cell lymphoma, acutemyeloid lymphoma, follicular lymphoma, chronic lymphocytic leukemia;breast, lung, endometrial, ovarian, gastric, cervical, and prostatecancer; glioblastoma; renal carcinoma; hepatocellular carcinoma; coloncarcinoma; neuroendocrine tumors; head and neck tumors; sarcomas; tumorsyndromes resulting directly or indirectly from genetic defects in PTEN(Phosphatase and tensin homologue deleted on chromosome 10), TSC1(Tuberous sclerosis 1), TSC2 (Tuberous sclerosis 2), NF1 (Neurofibromin1), AMPK (AMP-dependent protein kinase STK11, serine/threonine kinase11), and LKB1, VHL (von Hippel-Lindau disease) and PKD1 (polycystin-1);Cowden's disease, Cowden syndrome, Cowden-like syndrome, Bannayan-Zonanasyndrome, Bannayan-Riley-Ruvalcaba syndrome, Lhermitte-Duclos disease,endometrial carcinoma, tuberous sclerosis complex,lymphangioleiomyomatosis, neurofibromatosis 1, Peutz-Jeghers syndrome,renal cell carcinoma, von Hippel-Lindau disease, Proteus syndrome, andpolycystic kidney disease; rheumatoid arthritis; rheumatoid spondylitis;osteoarthritis; gout; asthma, bronchitis; allergic rhinitis; chronicobstructive pulmonary disease; cystic fibrosis; inflammatory boweldisease; irritable bowel syndrome; mucous colitis; ulcerative colitis;Crohn's disease; Huntington's disease; gastritis; esophagitis;hepatitis; pancreatitis; nephritis; multiple sclerosis; lupuserythematosus; atherosclerosis; restenosis following angioplasty; leftventricular hypertrophy; myocardial infarction; stroke; ischemic damagesof heart, lung, gut, kidney, liver, pancreas, spleen and brain; acute orchronic organ transplant rejection; preservation of the organ fortransplantation; organ failure or loss of limb (e.g., including, but notlimited to, that resulting from ischemia-reperfusion injury, trauma,gross bodily injury, car accident, crush injury or transplant failure);graft versus host disease; endotoxin shock; multiple organ failure;psoriasis; burn from exposure to fire, chemicals or radiation; eczema;dermatitis; skin graft; ischemia; ischemic conditions associated withsurgery or traumatic injury (e.g., vehicle accident, gunshot wound orlimb crush); epilepsy; Alzheimer's disease; Parkinson's disease;immunological response to bacterial or viral infection; cachexia;angiogenic and proliferative diseases, including retinitis pigmentosa,solid tumors, and cancers of a variety of tissues such as colon, rectum,prostate, liver, lung, bronchus, pancreas, brain, head, neck, stomach,skin, kidney, cervix, blood, larynx, esophagus, mouth, pharynx, urinarybladder, ovary or uterine.

Provided herein are methods for treating or preventing a solid tumor,non-Hodgkin lymphoma or multiple myeloma, comprising administering aneffective amount of the solid form of Compound A (e.g., Form A, Form B,Form C, Form D, Form E or amorphous), an isotopologue of Compound A, ametabolite of Compound A (e.g, O-desmethyl Compound A) or apharmaceutical composition provided herein, to a subject having a solidtumor, non-Hodgkin lymphoma or multiple myeloma. In one embodiment, thesolid tumor, non-Hodgkin lymphoma or multiple myeloma, is rapamycinresistant.

In one embodiment, the non-Hodgkin lymphoma is diffuse large B-celllymphoma (DLBCL), follicular lymphoma (FL), acute myeloid leukemia(AML), mantle cell lymphoma (MCL), or ALK⁺ anaplastic large celllymphoma. In one embodiment, the non-Hodgkin lymphoma is advanced solidnon-Hodgkin lymphoma.

In one embodiment, the solid tumor is a neuroendocrine tumor. In certainembodiments, the neuroendocrine tumor is a neuroendocrine tumor of gutorigin. In certain embodiments, the neuroendocrine tumor is ofnon-pancreatic origin. In certain embodiments, the neuroendocrine tumoris non-pancreatic of gut origin. In certain embodiments, theneuroendocrine tumor is of unknown primary origin. In some embodiments,the neuroendocrine tumor is of non-gut origin, for example a bronchialneuroendocrine tumor, or a neuroendocrine tumor with origin in an organabove the diaphragm, for example, a laryngeal neuroendocrine tumor, apharyngeal neuroendocrine tumor, or a thyroid neuroendocrine tumor. Incertain embodiments, the neuroendocrine tumor is a symptomatic endocrineproducing tumor or a nonfunctional tumor. In certain embodiments, theneuroendocrine tumor is locally unresectable, metastatic moderate, welldifferentiated, low (grade 1) or intermediate (grade 2).

In one embodiment, the solid tumor is non-small cell lung cancer(NSCLC).

In another embodiments the solid tumor is glioblastoma multiforme (GBM).

In another embodiment, the solid tumor is hepatocellular carcinoma(HCC).

In another embodiment, the solid tumor is breast cancer. In oneembodiment, the breast cancer is estrogen receptor positive (ER+,ER+/Her2− or ER+/Her2+). In one embodiment, the breast cancer isestrogen receptor negative (ER−/Her2+). In one embodiment, the breastcancer is triple negative (TN) (breast cancer that does not express thegenes and/or protein corresponding to the estrogen receptor (ER),progesterone receptor (PR), and that does not overexpress the Her2/neuprotein).

In another embodiment, the solid tumor is colorectal cancer.

In another embodiment, the solid tumor is salivary cancer.

In another embodiment, the solid tumor is pancreatic cancer.

In another embodiment, the solid tumor is adenocystic cancer.

In another embodiment, the solid tumor is adrenal cancer.

In another embodiment, the solid tumor is esophageal cancer.

In another embodiment, the solid tumor is renal cancer.

In another embodiment, the solid tumor is leiomyosarcoma.

In another embodiment, the solid tumor is paraganglioma.

In one embodiment, the solid tumor is an advanced solid tumor.

In one embodiment, the advanced solid tumor is a neuroendocrine tumor.In certain embodiments, the neuroendocrine tumor is a neuroendocrinetumor of gut origin. In certain embodiments, the neuroendocrine tumor isof non-pancreatic origin. In certain embodiments, the neuroendocrinetumor is non-pancreatic of gut origin. In certain embodiments, theneuroendocrine tumor is of unknown primary origin. In some embodiments,the neuroendocrine tumor is of non-gut origin, for example a bronchialneuroendocrine tumor, or a neuroendocrine tumor with origin in an organabove the diaphragm, for example, a laryngeal neuroendocrine tumor, apharyngeal neuroendocrine tumor, or a thyroid neuroendocrine tumor. Incertain embodiments, the neuroendocrine tumor is a symptomatic endocrineproducing tumor or a nonfunctional tumor. In certain embodiments, theneuroendocrine tumor is locally unresectable, metastatic moderate, welldifferentiated, low (grade 1) or intermediate (grade 2).

In one embodiment, the advanced solid tumor is non-small cell lungcancer (NSCLC).

In another embodiments the advanced solid tumor is glioblastomamultiforme (GBM).

In another embodiment, the advanced solid tumor is hepatocellularcarcinoma (HCC).

In another embodiment, the advanced solid tumor is breast cancer. In oneembodiment, the advanced solid tumor is estrogen receptor positive (ER+,ER+/Her2− or ER+/Her2+) breast cancer. In one embodiment, the advancedsolid tumor is ER+/Her2− breast cancer. In one embodiment, the advancedsolid tumor is ER+/Her2+ breast cancer. In one embodiment, the advancedsolid tumor is ER−/Her2+ breast cancer. In one embodiment, the advancedsolid tumor is triple negative (TN) breast cancer.

In another embodiment, the advanced solid tumor is colorectal cancer.

In another embodiment, the advanced solid tumor is salivary cancer.

In another embodiment, the advanced solid tumor is pancreatic cancer.

In another embodiment, the advanced solid tumor is adenocystic cancer.

In another embodiment, the advanced solid tumor is adrenal cancer.

In another embodiment, the advanced solid tumor is esophageal cancer.

In another embodiment, the advanced solid tumor is renal cancer.

In another embodiment, the advanced solid tumor is leiomyosarcoma.

In another embodiment, the advanced solid tumor is or paraganglioma.

In one embodiment, the non-Hodgkin lymphoma is diffuse large B-celllymphoma (DLBCL).

In one embodiment, provided herein are methods for achieving a ResponseEvaluation Criteria in Solid Tumors (RECIST 1.1) (see Eisenhauer E. A.,Therasse P., Bogaerts J., et al. New response evaluation criteria insolid tumours: Revised RECIST guideline (version 1.1). European J.Cancer; 2009; (45) 228-247) of complete response, partial response orstable disease in a patient comprising administering an effective amountof the solid form of Compound A (e.g., Form A, Form B, Form C, Form D,or Form E) or a pharmaceutical composition comprising the solid form ofCompound A (e.g., Form A, Form B, Form C, Form D, or Form E) providedherein, to a subject having a solid tumor, such as an advanced solidtumor.

In one embodiment, provided herein are methods for preventing ordelaying a Response Evaluation Criteria in Solid Tumors (RECIST 1.1) ofprogressive disease in a subject, comprising administering an effectiveamount of the solid form of Compound A (e.g., Form A, Form B, Form C,Form D, Form E or amorphous) or a pharmaceutical composition comprisingthe solid form of Compound A (e.g., Form A, Form B, Form C, Form D, FormE or amorphous) provided herein, to a subject having a solid tumor, suchas an advanced solid tumor. In one embodiment the prevention or delayingof progressive disease is characterized or achieved by a change inoverall size of the target lesions, of for example, between −30% and+20% compared to pre-treatment. In another embodiment, the change insize of the target lesions is a reduction in overall size of more than30%, for example, more than 50% reduction in target lesion size comparedto pre-treatment. In another, the prevention is characterized orachieved by a reduction in size or a delay in progression of non-targetlesions compared to pre-treatment. In one embodiment, the prevention isachieved or characterized by a reduction in the number of target lesionscompared to pre-treatment. In another, the prevention is achieved orcharacterized by a reduction in the number or quality of non-targetlesions compared to pre-treatment. In one embodiment, the prevention isachieved or characterized by the absence or the disappearance of targetlesions compared to pre-treatment. In another, the prevention isachieved or characterized by the absence or the disappearance ofnon-target lesions compared to pre-treatment. In another embodiment, theprevention is achieved or characterized by the prevention of new lesionscompared to pre-treatment. In yet another embodiment, the prevention isachieved or characterized by the prevention of clinical signs orsymptoms of disease progression compared to pre-treatment, such ascancer-related cachexia or increased pain.

In certain embodiments, provided herein are methods for decreasing thesize of target lesions in a subject compared to pre-treatment,comprising administering an effective amount of the solid form ofCompound A (e.g., Form A, Form B, Form C, Form D, Form E or amorphous),an isotopologue of Compound A, a metabolite of Compound A (e.g,O-desmethyl Compound A) or a pharmaceutical composition provided herein,to a subject having a solid tumor, such as an advanced solid tumor.

In certain embodiments, provided herein are methods for decreasing thesize of a non-target lesion in a subject compared to pre-treatment,comprising administering an effective amount of the solid form ofCompound A (e.g., Form A, Form B, Form C, Form D, Form E or amorphous),an isotopologue of Compound A, a metabolite of Compound A (e.g,O-desmethyl Compound A) or a pharmaceutical composition provided herein,to a subject having a solid tumor, such as an advanced solid tumor.

In certain embodiments, provided herein are methods for achieving areduction in the number of target lesions in a subject compared topre-treatment, comprising administering an effective amount of the solidform of Compound A (e.g., Form A, Form B, Form C, Form D, Form E oramorphous), an isotopologue of Compound A, a metabolite of Compound A(e.g, O-desmethyl Compound A) or a pharmaceutical composition providedherein, to a subject having a solid tumor, such as an advanced solidtumor.

In certain embodiments, provided herein are methods for achieving areduction in the number of non-target lesions in a subject compared topre-treatment, comprising administering an effective amount of the solidform of Compound A (e.g., Form A, Form B, Form C, Form D, Form E oramorphous), an isotopologue of Compound A, a metabolite of Compound A(e.g, O-desmethyl Compound A) or a pharmaceutical composition providedherein, to a subject having a solid tumor, such as an advanced solidtumor.

In certain embodiments, provided herein are methods for achieving anabsence of all target lesions in a subject, comprising administering aneffective amount of the solid form of Compound A (e.g., Form A, Form B,Form C, Form D, Form E or amorphous), an isotopologue of Compound A, ametabolite of Compound A (e.g, O-desmethyl Compound A) or apharmaceutical composition provided herein, to a subject having a solidtumor, such as an advanced solid tumor.

In certain embodiments, provided herein are methods for achieving anabsence of all non-target lesions in a subject, comprising administeringan effective amount of the solid form of Compound A (e.g., Form A, FormB, Form C, Form D, Form E or amorphous), an isotopologue of Compound A,a metabolite of Compound A (e.g, O-desmethyl Compound A) or apharmaceutical composition provided herein, to a subject having a solidtumor, such as an advanced solid tumor.

A method of treating a solid tumor, such as an advanced solid tumor, themethod comprising administering an effective amount of the solid form ofCompound A (e.g., Form A, Form B, Form C, Form D, Form E or amorphous),an isotopologue of Compound A, a metabolite of Compound A (e.g,O-desmethyl Compound A) or a pharmaceutical composition provided herein,to a subject having a solid tumor, such as an advanced solid tumor,wherein the treatment results in a complete response, partial responseor stable disease, as determined by Response Evaluation Criteria inSolid Tumors (RECIST 1.1).

A method of treating a solid tumor, such as an advanced solid tumor, themethod comprising administering an effective amount of the solid form ofCompound A (e.g., Form A, Form B, Form C, Form D, Form E or amorphous),an isotopologue of Compound A, a metabolite of Compound A (e.g,O-desmethyl Compound A) or a pharmaceutical composition provided herein,to a subject having a solid tumor, such as an advanced solid tumor,wherein the treatment results in a reduction in target lesion size, areduction in non-target lesion size and/or the absence of new targetand/or non-target lesions, compared to pre-treatment.

A method of treating a solid tumor, such as an advanced solid tumor, themethod comprising administering an effective amount of the solid form ofCompound A (e.g., Form A, Form B, Form C, Form D, Form E or amorphous),an isotopologue of Compound A, a metabolite of Compound A (e.g,O-desmethyl Compound A) or a pharmaceutical composition provided herein,to a subject having a solid tumor, such as an advanced solid tumor,wherein the treatment results in prevention or retarding of clinicalprogression, such as cancer-related cachexia or increased pain.

In another embodiment, provided herein are methods for improving theInternational Workshop Criteria (IWC) for NHL (see Cheson B D, PfistnerB, Juweid, M E, et. al. Revised Response Criteria for MalignantLymphoma. J. Clin. Oncol: 2007: (25) 579-586.) of a subject comprisingadministering an effective amount of the solid form of Compound A (e.g.,Form A, Form B, Form C, Form D, Form E or amorphous), an isotopologue ofCompound A, a metabolite of Compound A (e.g, O-desmethyl Compound A) ora pharmaceutical composition provided herein, to a subject havingnon-Hodgkin lymphoma. In another embodiment, provided herein are methodsto increase Progression Free Survival rates, as determined byKaplan-Meier estimates. In one embodiment, the treatment results in acomplete remission, partial remission or stable disease, as determinedby the International Workshop Criteria (IWC) for NHL. In anotherembodiment, the treatment results in an increase in overall survival,progression-free survival, event-free survival, time to progression,disease-free survival or lymphoma-free survival.

In another embodiment, provided herein are methods for inducing atherapeutic response characterized with the International UniformResponse Criteria for Multiple Myeloma (IURC) (see Durie BGM, HarousseauJ-L, Miguel J S, et al. International uniform response criteria formultiple myeloma. Leukemia, 2006; (10) 10: 1-7) of a subject comprisingadministering an effective amount of the solid form of Compound A (e.g.,Form A, Form B, Form C, Form D, Form E or amorphous), an isotopologue ofCompound A, a metabolite of Compound A (e.g, O-desmethyl Compound A) ora pharmaceutical composition provided herein, to a subject havingmultiple myeloma. In one embodiment, the treatment results in astringent complete response, complete response, or very good partialresponse, as determined by the the International Uniform ResponseCriteria for Multiple Myeloma (IURC). In another embodiment, thetreatment results in an increase in overall survival, progression-freesurvival, event-free survival, time to progression, or disease-freesurvival.

In another embodiment, provided herein are methods for inducing atherapeutic response assessed with Response Assessment forNeuro-Oncology (RANO) Working Group for GBM (see Wen P., Macdonald, DR., Reardon, D A., et al. Updated response assessment criteria forhighgrade gliomas: Response assessment in neuro-oncology working group.J. Clin. Oncol. 2010; 28: 1963-1972) of a subject comprisingadministering an effective amount of the solid form of Compound A (e.g.,Form A, Form B, Form C, Form D, Form E or amorphous), an isotopologue ofCompound A, a metabolite of Compound A (e.g, O-desmethyl Compound A) ora pharmaceutical composition provided herein, to a subject havinggliobastoma multiforme.

In another embodiment, provided herein are methods for improving theEastern Cooperative Oncology Group Performance Status (ECOG) of asubject comprising administering an effective amount of the solid formof Compound A (e.g., Form A, Form B, Form C, Form D, Form E oramorphous), an isotopologue of Compound A, a metabolite of Compound A(e.g, O-desmethyl Compound A) or a pharmaceutical composition providedherein, to a subject having a tumor, such as an advanced solid tumor.

In another embodiment, provided herein are methods for inducing atherapeutic response assessed by Positron Emission Tomography (PET)outcome of a subject comprising administering an effective amount of thesolid form of Compound A (e.g., Form A, Form B, Form C, Form D, Form Eor amorphous), an isotopologue of Compound A, a metabolite of Compound A(e.g, O-desmethyl Compound A) or a pharmaceutical composition providedherein, to a subject having a tumor, such as an advanced solid tumor. Incertain embodiments, provided herein are methods for treating a solidtumor, such as an advanced solid tumor, the methods comprisingadministering an effective amount of a TOR kinase inhibitor to a patienthaving a solid tumor, such as an advanced solid tumor, wherein thetreatment results in a reduction in tumor metabolic activity, forexample, as measured by PET imaging.

In another embodiment, provided herein are methods for inducing atherapeutic response assessed by a reduction in carcinoidsyndrome-related symptoms, such as diarrhea and/or flushing, and/or areduction in endocrine hormone markers, such as chromogranin, gastrin,serotonin, and/or glucagon.

In one embodiment, provided herein are methods for inhibitingphosphorylation of S6RP, 4E-BP1 and/or AKT in a subject having a solidtumor (for example, a neuroendocrine tumor, non-small cell lung cancer,glioblastoma multiforme, hepatocellular carcinoma, breast cancer,colorectal cancer, salivary cancer, pancreatic cancer, adenocysticcancer, adrenal cancer, esophageal cancer, renal cancer, leiomyosarcoma,or paraganglioma), non-Hodgkin lymphoma or multiple myeloma, comprisingadministering an effective amount of the solid form of Compound A (e.g.,Form A, Form B, Form C, Form D, Form E or amorphous), an isotopologue ofCompound A, a metabolite of Compound A (e.g, O-desmethyl Compound A) ora pharmaceutical composition provided herein to said subject. In somesuch embodiments, the inhibition of phosphorylation is assessed in abiological sample of the subject, such as in circulating blood and/ortumor cells, skin biopsies and/or tumor biopsies or aspirate. In suchembodiments, the amount of inhibition of phosphorylation is assessed bycomparison of the amount of phospho-S6RP, 4E-BP1 and/or AKT before andafter administration of the solid form of Compound A (e.g., Form A, FormB, Form C, Form D, Form E or amorphous), an isotopologue of Compound A,a metabolite of Compound A (e.g, O-desmethyl Compound A) or apharmaceutical composition provided herein. In certain embodiments,provided herein are methods for measuring inhibition of phosphorylationof S6RP, 4E-BP1 or AKT in a subject having a solid tumor (for example, aneuroendocrine tumor, non-small cell lung cancer, glioblastomamultiforme, hepatocellular carcinoma, breast cancer, colorectal cancer,salivary cancer, pancreatic cancer, adenocystic cancer, adrenal cancer,esophageal cancer, renal cancer, leiomyosarcoma, or paraganglioma),non-Hodgkin lymphoma or multiple myeloma, comprising administering aneffective amount of the solid form of Compound A (e.g., Form A, Form B,Form C, Form D, Form E or amorphous), an isotopologue of Compound A, ametabolite of Compound A (e.g, O-desmethyl Compound A) or apharmaceutical composition provided herein to said subject, measuringthe amount of phosphorylated S6RP, 4E-BP1 and/or AKT in said subject,and comparing said amount of phosphorylated S6RP, 4E-BP1 and/or AKT tothat of said subject prior to administration of an effective amount ofthe solid form of Compound A (e.g., Form A, Form B, Form C, Form D, FormE or amorphous), an isotopologue of Compound A, a metabolite of CompoundA (e.g, O-desmethyl Compound A) or a pharmaceutical composition providedherein. In some embodiments, the inhibition of phosphorylation of S6RP,4E-BP1 and/or AKT is assessed in B-cells, T-cells and/or monocytes.

In certain embodiments, provided herein are methods for inhibitingphosphorylation of S6RP, 4E-BP1 and/or AKT in a biological sample of asubject having a solid tumor (for example, a neuroendocrine tumor,non-small cell lung cancer, glioblastoma multiforme, hepatocellularcarcinoma, breast cancer, colorectal cancer, salivary cancer, pancreaticcancer, adenocystic cancer, adrenal cancer, esophageal cancer, renalcancer, leiomyosarcoma, or paraganglioma), non-Hodgkin lymphoma ormultiple myeloma, comprising administering an effective amount of thesolid form of Compound A (e.g., Form A, Form B, Form C, Form D, Form Eor amorphous), an isotopologue of Compound A, a metabolite of Compound A(e.g, O-desmethyl Compound A) or a pharmaceutical composition providedherein to said subject and comparing the amount of phosphorylated S6RP,4E-BP1 and/or AKT in a biological sample of a subject obtained prior toand after administration of said solid form of Compound A (e.g., Form A,Form B, Form C, Form D, Form E or amorphous), isotopologue of CompoundA, metabolite of Compound A (e.g, O-desmethyl Compound A) orpharmaceutical composition provided herein, wherein less phosphorylatedS6RP, 4E-BP1 and/or AKT in said biological sample obtained afteradministration of said solid form of Compound A (e.g., Form A, Form B,Form C, Form D, Form E or amorphous), isotopologue of Compound A,metabolite of Compound A (e.g, O-desmethyl Compound A) or apharmaceutical composition provided herein relative to the amount ofphosphorylated S6RP, 4E-BP1 and/or AKT in said biological sampleobtained prior to administration of said solid form of Compound A (e.g.,Form A, Form B, Form C, Form D, Form E or amorphous), isotopologue ofCompound A, metabolite of Compound A (e.g, O-desmethyl Compound A) orpharmaceutical composition provided herein indicates inhibition. In someembodiments, the inhibition of phosphorylation of S6RP, 4E-BP1 and/orAKT is assessed in B-cells, T-cells and/or monocytes.

In one embodiment, provided herein are methods for inhibitingDNA-dependent protein kinase (DNA-PK) activity in a subject having asolid tumor (for example, a neuroendocrine tumor, non-small cell lungcancer, glioblastoma multiforme, hepatocellular carcinoma, breastcancer, colorectal cancer, salivary cancer, pancreatic cancer,adenocystic cancer, adrenal cancer, esophageal cancer, renal cancer,leiomyosarcoma, or paraganglioma), non-Hodgkin lymphoma or multiplemyeloma, comprising administering an effective amount of the solid formof Compound A (e.g., Form A, Form B, Form C, Form D, Form E oramorphous), an isotopologue of Compound A, a metabolite of Compound A(e.g, O-desmethyl Compound A) or a pharmaceutical composition providedherein to said subject. In some embodiments, DNA-PK inhibition isassessed in the skin of the subject having a solid tumor (for example, aneuroendocrine tumor, non-small cell lung cancer, glioblastomamultiforme, hepatocellular carcinoma, breast cancer, colorectal cancer,salivary cancer, pancreatic cancer, adenocystic cancer, adrenal cancer,esophageal cancer, renal cancer, leiomyosarcoma, or paraganglioma),non-Hodgkin lymphoma or multiple myeloma, in one example in a UVlight-irradiated skin sample of said subject. In another embodiment,DNA-PK inhibition is assessed in a tumor biopsy or aspirate of a subjecthaving a solid tumor (for example, a neuroendocrine tumor, non-smallcell lung cancer, glioblastoma multiforme, hepatocellular carcinoma,breast cancer, colorectal cancer, salivary cancer, pancreatic cancer,adenocystic cancer, adrenal cancer, esophageal cancer, renal cancer,leiomyosarcoma, or paraganglioma), non-Hodgkin lymphoma or multiplemyeloma. In one embodiment, inhibition is assessed by measuring theamount of phosphorylated DNA-PK S2056 (also known as pDNA-PK S2056)before and after administration of the solid form of Compound A (e.g.,Form A, Form B, Form C, Form D, Form E or amorphous), an isotopologue ofCompound A, a metabolite of Compound A (e.g, O-desmethyl Compound A) ora pharmaceutical composition provided herein. In certain embodiments,provided herein are methods for measuring inhibition of phosphorylationof DNA-PK S2056 in a skin sample of a subject having a solid tumor (forexample, a neuroendocrine tumor, non-small cell lung cancer,glioblastoma multiforme, hepatocellular carcinoma, breast cancer,colorectal cancer, salivary cancer, pancreatic cancer, adenocysticcancer, adrenal cancer, esophageal cancer, renal cancer, leiomyosarcoma,or paraganglioma), non-Hodgkin lymphoma or multiple myeloma, comprisingadministering an effective amount of the solid form of Compound A (e.g.,Form A, Form B, Form C, Form D, Form E or amorphous), an isotopologue ofCompound A, a metabolite of Compound A (e.g, O-desmethyl Compound A) ora pharmaceutical composition provided herein to said subject, measuringthe amount of phosphorylated DNA-PK S2056 present in the skin sample andcomparing said amount of phosphorylated DNA-PK S2056 to that in a skinsample from said subject prior to administration of an effective amountof the solid form of Compound A (e.g., Form A, Form B, Form C, Form D,Form E or amorphous), an isotopologue of Compound A, a metabolite ofCompound A (e.g, O-desmethyl Compound A) or a pharmaceutical compositionprovided herein. In one embodiment, the skin sample is irradiated withUV light.

In certain embodiments, provided herein are methods for inhibitingDNA-dependent protein kinase (DNA-PK) activity in a skin sample of asubject having a solid tumor (for example, a neuroendocrine tumor,non-small cell lung cancer, glioblastoma multiforme, hepatocellularcarcinoma, breast cancer, colorectal cancer, salivary cancer, pancreaticcancer, adenocystic cancer, adrenal cancer, esophageal cancer, renalcancer, leiomyosarcoma, or paraganglioma), non-Hodgkin lymphoma ormultiple myeloma, comprising administering an effective amount of thesolid form of Compound A (e.g., Form A, Form B, Form C, Form D, Form Eor amorphous), an isotopologue of Compound A, a metabolite of Compound A(e.g, O-desmethyl Compound A) or a pharmaceutical composition providedhereinto said subject and comparing the amount of phosphorylated DNA-PKin a biological sample of a subject obtained prior to and afteradministration of said solid form of Compound A (e.g., Form A, Form B,Form C, Form D, Form E or amorphous), isotopologue of Compound A,metabolite of Compound A (e.g, O-desmethyl Compound A) or pharmaceuticalcomposition provided herein, wherein less phosphorylated DNA-PK in saidbiological sample obtained after administration of said solid form ofCompound A (e.g., Form A, Form B, Form C, Form D, Form E or amorphous),isotopologue of Compound A, metabolite of Compound A (e.g, O-desmethylCompound A) or pharmaceutical composition provided herein relative tothe amount of phosphorylated DNA-PK in said biological sample obtainedprior to administration of said solid form of Compound A (e.g., Form A,Form B, Form C, Form D, Form E or amorphous), isotopologue of CompoundA, metabolite of Compound A (e.g, O-desmethyl Compound A) orpharmaceutical composition provided herein indicates inhibition.

The solid form of Compound A (e.g., Form A, Form B, Form C, Form D, FormE or amorphous), isotopologues of Compound A, metabolites of Compound A(e.g, O-desmethyl Compound A) and pharmaceutical compositions providedherein can be combined with radiation therapy or surgery. In certainembodiments, the solid form of Compound A (e.g., Form A, Form B, Form C,Form D, Form E or amorphous), isotopologues of Compound A, metabolitesof Compound A (e.g, O-desmethyl Compound A) and pharmaceuticalcompositions provided herein are administered to subject who isundergoing radiation therapy, has previously undergone radiation therapyor will be undergoing radiation therapy. In certain embodiments, thesolid form of Compound A (e.g., Form A, Form B, Form C, Form D, Form Eor amorphous), isotopologues of Compound A, metabolites of Compound A(e.g, O-desmethyl Compound A) and pharmaceutical compositions providedherein are administered to a subject who has undergone tumor removalsurgery (e.g., surgery to remove a GBM tumor).

Further provided herein are methods for treating subjects who have beenpreviously treated for a solid tumor (for example, a neuroendocrinetumor, non-small cell lung cancer, glioblastoma multiforme,hepatocellular carcinoma, breast cancer, colorectal cancer, salivarycancer, pancreatic cancer, adenocystic cancer, adrenal cancer,esophageal cancer, renal cancer, leiomyosarcoma, or paraganglioma),non-Hodgkin lymphoma or multiple myeloma, but are non-responsive tostandard therapies, as well as those who have not previously beentreated. Further provided herein are methods for treating subjects whohave undergone surgery in an attempt to treat the condition at issue, aswell as those who have not. Because subjects with a solid tumor (forexample, a neuroendocrine tumor, non-small cell lung cancer,glioblastoma multiforme, hepatocellular carcinoma, breast cancer,colorectal cancer, salivary cancer, pancreatic cancer, adenocysticcancer, adrenal cancer, esophageal cancer, renal cancer, leiomyosarcoma,or paraganglioma), non-Hodgkin lymphoma or multiple myeloma haveheterogenous clinical manifestations and varying clinical outcomes, thetreatment given to a subject may vary, depending on his/her prognosis.

In certain embodiments, the pharmaceutical compositions provided hereincomprising Compound A can be used for the treatment or prevention of adisease disclosed in U.S. Pat. Appl. Publ. No. 2010/0216781 (see, e.g.,paragraphs [0415]-[0437]), the disclosure of which is incorporatedherein by reference in its entirety.

Further provided herein are methods for achieving certainpharmacokinetic (PK) parameters with respect to Compound A in a subject,comprising administering a pharmaceutical composition provided herein tosaid subject. In certain embodiments, provided herein are methods forachieving a PK parameter set forth in the examples provided herein withrespect to Compound A in a subject, comprising administering apharmaceutical composition provided herein to said subject. In certainembodiments, the methods for achieving a PK parameter described hereinfurther comprise measuring the amount of Compound A in a biologicalsample (e.g., urine, blood, serum or plasma) of a subject afteradministration of Compound A.

In certain embodiments, provided herein are methods for achieving a Tmaxof about 0.5 to about 2 hours of Compound A in a subject, comprisingadministering a pharmaceutical composition provided herein to saidsubject. In specific embodiments, provided herein are methods forachieving a T_(max) of about 1 hour, about 1.5 hours or about 2 hours ofCompound A in a subject, comprising administering a pharmaceuticalcomposition provided herein to said subject.

In certain embodiments, provided herein are methods for achieving at_(1/2) of about 4 to about 8 hours of Compound A in a subject,comprising administering a pharmaceutical composition provided herein tosaid subject. In specific embodiments, provided herein are methods forachieving a t_(1/2) of about 4 hours, about 4.5 hours, about 5 hours,about 5.5 hours, about 6 hours, about 6.5 hours, about 7 hours, about7.5 hours or about 8 hours of Compound A in a subject, comprisingadministering a pharmaceutical composition provided herein to saidsubject.

In certain embodiments, provided herein are methods for achieving aC_(max) of about 150 to about 500 ng/mL of Compound A in a subject,comprising administering a pharmaceutical composition provided herein tosaid subject. In specific embodiments, provided herein are methods forachieving a C_(max) of about 150 ng/mL, about 175 ng/mL, about 200ng/mL, about 225 ng/mL, about 250 ng/mL, about 275 ng/mL, about 300ng/mL, about 325 ng/mL, about 350 ng/mL, about 375 ng/mL, about 400ng/mL, about 425 ng/mL, about 450 ng/mL, about 475 ng/mL or about 500ng/mL of Compound A in a subject, comprising administering apharmaceutical composition provided herein to said subject. In oneembodiment, provided herein are methods for achieving a steady stateCmax of about 485 ng/mL of Compound A in a subject, comprisingadministering a pharmaceutical composition provided herein to saidsubject.

In certain embodiments, provided herein are methods for achieving anAUC₀₋₂₄ of about 900 to about 2500 ng*h/mL of Compound A in a subject,comprising administering a pharmaceutical composition provided herein tosaid subject. In specific embodiments, provided herein are methods forachieving an AUC₀₋₂₄ of about 900 ng*hr/mL, about 950 ng*hr/mL, about1000 ng*hr/mL, about 1050 ng*hr/mL, about 1100 ng*hr/mL, about 1150ng*hr/mL, about 1200 ng*hr/mL, about 1250 ng*hr/mL, about 1300 ng*hr/mL,about 1350 ng*hr/mL, about 1400 ng*hr/mL, about 1450 ng*hr/mL, about1500 ng*hr/mL, about 1550 ng*hr/mL, about 1600 ng*hr/mL, about 1650ng*hr/mL, about 1700 ng*hr/mL, about 1750 ng*hr/mL, about 1800 ng*hr/mL,about 1850 ng*hr/mL, about 1900 ng*hr/mL, about 1950 ng*hr/mL, about2000 ng*hr/mL, about 2050 ng*hr/mL, about 2100 ng*hr/mL, about 2150ng*hr/mL, about 2200 ng*hr/mL, about 2250 ng*hr/mL, about 2300 ng*hr/mL,about 2350 ng*hr/mL, about 2400 ng*hr/mL, about 2450 ng*hr/mL or about2500 ng*hr/mL of Compound A in a subject, comprising administering apharmaceutical composition provided herein to said subject.

In certain embodiments, provided herein are methods for achieving anAUC_(∞) of about 900 to about 1100 ng*hr/mL of Compound A in a subject,comprising administering a pharmaceutical composition provided herein tosaid subject. In specific embodiments, provided herein are methods forachieving an AUC_(∞) of about 900 ng*hr/mL, about 950 ng*hr/mL, about1000 ng*hr/mL, about 1050 ng*hr/mL or about 1000 ng*hr/mL of Compound Ain a subject, comprising administering a pharmaceutical compositionprovided herein to said subject.

In certain embodiments, provided herein are methods for achieving a CL/Fof about 19 to about 22 L/hr of Compound A in a subject, comprisingadministering a pharmaceutical composition provided herein to saidsubject. In specific embodiments, provided herein are methods forachieving a CL/F of about 19 L/hr, about 19.5 L/hr, about 20 L/hr, about20.5 L/hr, about 21 L/hr, about 21.5 L/hr or about 22 L/hr of Compound Ain a subject, comprising administering a pharmaceutical compositionprovided herein to said subject.

In certain embodiments, provided herein are methods for achieving a Vz/Fof about 150 to about 180 L of Compound A in a subject, comprisingadministering a pharmaceutical composition provided herein to saidsubject. In specific embodiments, provided herein are methods forachieving a Vz/F of about 150 L, about 155 L, about 160 L, about 165 L,about 170 L, about 175 L or about 180 L of Compound A in a subject,comprising administering a pharmaceutical composition provided herein tosaid subject.

In certain embodiments, the methods of use and pharmaceuticalcompositions provided herein comprise in vivo production of a metaboliteof Compound A.

In certain embodiments, the methods of use and pharmaceuticalcompositions provided herein comprise in vivo production of a metaboliteof Compound A in a subject, wherein the metabolite has one or more ofthe pharmacokinetic parameters selected from a C_(max) of about 100 toabout 200 ng/mL (e.g., 143 ng/mL), a T_(max) of about 7 to about 9 hours(e.g., 8 hours), an AUC₀₋₂₄ of about 2500 to about 3000 ng*h/mL (e.g.,2744 ng*h/mL), an AUC_(0-∞) of about 7750 to about 8250 ng*h/mL (e.g.,7948 ng*h/mL) and a t_(1/2) of about 30 to about 40 hours (e.g., 35hours) on day 1 of administration of about 7.5 mg of Compound A or apharmaceutical composition thereof to said subject or wherein themetabolite has one or more of the pharmacokinetic parameters selectedfrom a C_(max) of about 300 to about 400 ng/mL (e.g., 363 ng/mL), aT_(max) of about 1 to about 3 hours (e.g., 2 hours), an AUC₀₋₂₄ of about6250 to about 6750 ng*h/mL (e.g., 6404 ng*h/mL), an AUC_(0-∞) of about42500 to about 47500 ng*h/mL (e.g., 45602 ng*h/mL) and a C_(trough) ofabout 200 to about 300 ng/mL (e.g., 267 ng/mL) on day 15 of once a dayadministration of about 7.5 mg of Compound A or a pharmaceuticalcomposition thereof to said subject.

In certain embodiments, the methods of use and pharmaceuticalcompositions provided herein comprise in vivo production of a metaboliteof Compound A in a subject, wherein the metabolite has one or more ofthe pharmacokinetic parameters selected from a C_(max) of about 250 toabout 350 ng/mL (e.g., 309 ng/mL), a T_(max) of about 1 to about 3 hours(e.g., 2 hours), an AUC₀₋₂₄ of about 3500 to about 4000 ng*h/mL (e.g.,3828 ng*h/mL), an AUC_(0-∞) of about 5500 to about 6000 ng*h/mL (e.g.,5821 ng*h/mL) and a t_(1/2) of about 10 to about 14 hours (e.g., 12hours) on day 1 of administration of about 15 mg of Compound A or apharmaceutical composition thereof to said subject or wherein themetabolite has one or more of the pharmacokinetic parameters selectedfrom a C_(max) of about 400 to about 500 ng/mL (e.g., 458 ng/mL), aT_(max) of about 2 to about 4 hours (e.g., 3 hours), an AUC₀₋₂₄ of about5500 to about 6000 ng*h/mL (e.g., 5677 ng*h/mL), an AUC_(0-∞) of about9500 to about 10000 ng*h/mL (e.g., 9753 ng*h/mL) and a C_(trough) ofabout 100 to about 200 ng/mL (e.g., 145 ng/mL) on day 15 of once a dayadministration of about 15 mg of Compound A or a pharmaceuticalcomposition thereof to said subject.

In certain embodiments, the methods of use and pharmaceuticalcompositions provided herein comprise in vivo production of a metaboliteof Compound A in a subject, wherein the metabolite has one or more ofthe pharmacokinetic parameters selected from a C_(max) of about 700 toabout 800 ng/mL (e.g., 776 ng/mL), a T_(max) of about 6 to about 8 hours(e.g., 7 hours), an AUC₀₋₂₄ of about 13000 to about 13500 ng*h/mL (e.g.,13288 ng*h/mL), an AUC_(0-∞) of about 25000 to about 30000 ng*h/mL(e.g., 27672 ng*h/mL) and a t_(1/2) of about 18 to about 24 hours (e.g.,21 hours) on day 1 of administration of about 30 mg of Compound A or apharmaceutical composition thereof to said subject or wherein themetabolite has one or more of the pharmacokinetic parameters selectedfrom a C_(max) of about 1600 to about 2000 ng/mL (e.g., 1768 ng/mL), aT_(max) of about 1 to about 3 hours (e.g., 2 hours), an AUC₀₋₂₄ of about27500 to about 32500 ng*h/mL (e.g., 29423 ng*h/mL), an AUC_(0-∞) ofabout 110000 to about 130000 ng*h/mL (e.g., 117697 ng*h/mL) and aC_(trough) of about 1000 to about 1200 ng/mL (e.g., 1102 ng/mL) on day15 of once a day administration of about 30 mg of Compound A or apharmaceutical composition thereof to said subject.

In certain embodiments, the methods of use and pharmaceuticalcompositions provided herein comprise in vivo production of a metaboliteof Compound A in a subject, wherein the metabolite has one or more ofthe pharmacokinetic parameters selected from a C_(max) of about 1100 toabout 1200 ng/mL (e.g., 1153 ng/mL), a T_(max) of about 2 to about 4hours (e.g., 3 hours), an AUC₀₋₂₄ of about 15500 to about 16000 ng*h/mL(e.g., 15854 ng*h/mL), an AUC_(0-∞) of about 25000 to about 30000ng*h/mL (e.g., 27274 ng*h/mL) and a t_(1/2) of about 14 to about 20hours (e.g., 17 hours) on day 1 of administration of about 45 mg ofCompound A or a pharmaceutical composition thereof to said subject orwherein the metabolite has one or more of the pharmacokinetic parametersselected from a C_(max) of about 2000 to about 2500 ng/mL (e.g., 2243ng/mL), a T_(max) of about 1 to about 3 hours (e.g., 2 hours), anAUC₀₋₂₄ of about 30000 to about 35000 ng*h/mL (e.g., 32705 ng*h/mL), anAUC_(0-∞) of about 75000 to about 80000 ng*h/mL (e.g., 77722 ng*h/mL)and a C_(trough) of about 1100 to about 1200 ng/mL (e.g., 1181 ng/mL) onday 15 of once a day administration of about 45 mg of Compound A or apharmaceutical composition thereof to said subject.

In certain embodiments, the methods of use and pharmaceuticalcompositions provided herein comprise in vivo production of a metaboliteof Compound A in a subject, wherein the metabolite has one or more ofthe pharmacokinetic parameters selected from a C_(max) of about 1400 toabout 1500 ng/mL (e.g., 1438 ng/mL), a T_(max) of about 4 to about 6hours (e.g., 5 hours), an AUC₀₋₂₄ of about 21000 to about 22000 ng*h/mL(e.g., 21454 ng*h/mL), an AUC_(0-∞) of about 35000 to about 40000ng*h/mL (e.g., 37490 ng*h/mL) and a t_(1/2) of about 12 to about 20hours (e.g., 16 hours) on day 1 of administration of about 60 mg ofCompound A or a pharmaceutical composition thereof to said subject orwherein the metabolite has one or more of the pharmacokinetic parametersselected from a C_(max) of about 2250 to about 2750 ng/mL (e.g., 2521ng/mL), a T_(max) of about 2 to about 4 hours (e.g., 3 hours), anAUC₀₋₂₄ of about 45000 to about 50000 ng*h/mL (e.g., 46852 ng*h/mL), anAUC_(0-∞) of about 135000 to about 145000 ng*h/mL (e.g., 138418 ng*h/mL)and a C_(trough) of about 1400 to about 1500 ng/mL (e.g., 1467 ng/mL) onday 15 of once a day administration of about 60 mg of Compound A or apharmaceutical composition thereof to said subject.

In certain embodiments, the methods of use and pharmaceuticalcompositions provided herein comprise in vivo production of a metaboliteof Compound A in a subject, wherein the metabolite has a T_(max) ofabout 2 to about 4 hours (e.g., 3 hours) upon administration of about 20mg of Compound A or a pharmaceutical composition thereof or about 45 mgof Compound A or a pharmaceutical composition thereof to said subject.

In certain embodiments, the methods of use and pharmaceuticalcompositions provided herein comprise in vivo production of a metaboliteof Compound A in a subject, wherein the metabolite has a C_(max) ofabout 450 to about 550 ng/mL (e.g., 503 ng/mL) upon administration ofabout 20 mg of Compound A or a pharmaceutical composition thereof or aC_(max) of about 1100 to about 1200 ng/mL (e.g., 1153 ng/mL) uponadministration of about 45 mg of Compound A or a pharmaceuticalcomposition thereof to said subject.

In certain embodiments, the methods of use and pharmaceuticalcompositions provided herein comprise in vivo production of a metaboliteof Compound A in a subject, wherein the metabolite has an AUC_(∞) ofabout 10000 to about 15000 ng/mL (e.g., 11928 ng*h/mL) uponadministration of about 20 mg of Compound A or a pharmaceuticalcomposition thereof or an AUC_(∞) of about 25000 to about 30000 ng/mL(e.g., 27274 ng*h/mL) upon administration of about 45 mg of Compound Aor a pharmaceutical composition thereof to said subject.

In certain embodiments, the methods of use and pharmaceuticalcompositions provided herein comprise in vivo production of a metaboliteof Compound A in a subject, wherein the metabolite has an AUC₀₋₂₄ ofabout 7000 to about 8000 ng/mL (e.g., 7484 ng*h/mL) upon administrationof about 20 mg of Compound A or a pharmaceutical composition thereof oran AUC₀₋₂₄ of about 12500 to about 17500 ng/mL (e.g., 15854 ng*h/mL)upon administration of about 45 mg of Compound A or a pharmaceuticalcomposition thereof to said subject.

In certain embodiments, the methods of use and pharmaceuticalcompositions provided herein comprise in vivo production of a metaboliteof Compound A in a subject, wherein the metabolite has a t_(1/2) ofabout 12 to about 16 hours (e.g., 14.3 hours) upon administration ofabout 20 mg of Compound A or a pharmaceutical composition thereof or at_(1/2) of about 12 to about 16 hours (e.g., 14.7 hours) uponadministration of about 45 mg of Compound A or a pharmaceuticalcomposition thereof to said subject.

In certain embodiments, the pharmacokinetic parameters in connectionwith the metabolite of Compound A produced via administration of 7.5 mg,15 mg, 30 mg, 45 mg and 60 mg of Compound A are obtained using theprotocol set forth in Section 5.2.1 (paragraphs [00497]-[00520]) of U.S.provisional application No. 61/653,436, filed May 31, 2012, which isincorporated by reference herein in its entirety.

In certain embodiments, the pharmacokinetic parameters in connectionwith the metabolite of Compound A produced via administration of 20 mgof Compound A were obtained using the protocol set forth in Section6.5.1, below.

In certain embodiments, the pharmacokinetic parameters set forth hereinare mean values obtained from multiple subjects.

In certain embodiments, the metabolite of Compound A is the O-desmethylmetabolite.

6. EXAMPLES

Chem-4D Draw (ChemInnovation Software, Inc., San Diego, Calif.),ChemDraw Ultra (Cambridgesoft, Cambridge, Mass.) or ACD/Name (AdvancedChemistry Development, Inc. Toronto, Ontario) was used to generate namesfor chemical structures.

The following abbreviations were used in descriptions and examples:

-   -   ACN Acetonitrile    -   Amphos Di-tert-butyl(4-dimethylaminophenyl)phosphine    -   BHT Butylated hydroxytoluene    -   Boc tert-Butoxycarbonyl    -   dba Dibenzylideneacetone    -   DCM Dichloromethane    -   DIBE Diisobutyl hexahydrophthalate    -   DIPEA N,N-Diisopropylethylamine    -   DIPE Diisopropyl ether    -   DME Dimethoxyethane    -   DMAP 4-Dimethylaminopyridine    -   DMSO Dimethylsulfoxide    -   dppf 1,1′-Bis(diphenylphosphino)ferrocene    -   DSC Differential scanning calorimetry    -   ESI Electrospray ionization    -   EtOAc Ethyl acetate    -   DVS Dynamic vapor sorption    -   HPLC High performance liquid chromatography    -   IPA Isopropyl alcohol    -   IPAc Isopropyl acetate    -   MeOAc Methyl acetate    -   MIBK Methyl isobutyl ketone    -   mp Melting point    -   MS Mass spectrometry    -   MTBE Methyl tert-butyl ether    -   NBS N-Bromosuccinimide    -   NMR Nuclear magnetic resonance    -   NMP N-methyl-2-pyrrolidinone    -   PEG Polyethylene glycol    -   PFL Protect from light    -   REF Refrigerated    -   RTmp or RT Room temperature    -   TEA Triethylamine    -   TFA Trifluoroacetic acid    -   TGA Thermogravimetric analysis    -   THF Tetrahydrofuran    -   TLC Thin layer chromatography    -   TMS Trimethylsilyl    -   XRPD X-ray powder diffraction

The following Examples are presented by way of illustration, notlimitation.

6.1 Solid Form Screen

6.1.1 Characterization Methodology

6.1.1.1 X-Ray Powder Diffraction (XRPD)

All of solid samples generated in the solid form screen were analyzed byXRPD. XRPD analysis was conducted on a Bruker AXS C2 GADDS or Bruker AXSD8 Advance X-ray powder diffractometer.

Certain X-Ray Powder Diffraction patterns were collected on a Bruker AXSC2 GADDS diffractometer using Cu Ka radiation (40 kV, 40 mA), automatedXYZ stage, laser video microscope for auto-sample positioning and aHiStar 2-dimensional area detector. X-ray optics consists of a singleGobel multilayer mirror coupled with a pinhole collimator of 0.3 mm. Aweekly performance check is carried out using a certified standard NIST1976 Corundum (flat plate). The beam divergence, i.e. the effective sizeof the X-ray beam on the sample, was approximately 4 mm. A θ-θcontinuous scan mode was employed with a sample—detector distance of 20cm which gives an effective 20 range of 3.2°-29.7°. Typically the samplewould be exposed to the X-ray beam for 120 seconds. The software usedfor data collection was GADDS for WNT 4.1.16 and the data were analyzedand presented using Diffrac Plus EVA vl11.0.0.2 or v13.0.0.2. Ambientconditions: Samples run under ambient conditions were prepared as flatplate specimens using powder as received without grinding. Approximately1-2 mg of the sample was lightly pressed on a glass slide to obtain aflat surface. Non-ambient conditions: Samples run under non-ambientconditions were mounted on a silicon wafer with heatconducting compound.The sample was then heated to the appropriate temperature at 20° C./minand subsequently held isothermally for 1 minute before data collectionwas initiated.

Certain X-Ray Powder Diffraction patterns were collected on a Bruker D8diffractometer using CuKa radiation (40 kV, 40 mA), θ-2 θ goniometer,and divergence of V4 and receiving slits, a Ge monochromator and aLynxeye detector. The instrument is performance checked using acertified Corundum standard (NIST 1976). The software used for datacollection was Diffrac Plus XRD Commander v2.5.0 and the data wereanalyzed and presented using Diffrac Plus EVA v11.0.0.2 or v13.0.0.2.Samples were run under ambient conditions as flat plate specimens usingpowder as received. The sample was gently packed into a cavity cut intopolished, zero-background (510) silicon wafer. The sample was rotated inits own plane during analysis. The details of the data collection are:Angular range: 2 to 42° 2θ; Step size: 0.05° 2 θ; Collection time: 0.5s/step.

6.1.1.2 Differential Scanning Calorimetry (DSC)

Modulated DSC data were collected on a TA Instruments Q2000 equippedwith a 50 position auto-sampler. The calibration for thermal capacitywas carried out using sapphire and the calibration for energy andtemperature was carried out using certified indium. Typically 3-1.5 mgof each sample, in a pin-holed aluminum pan, was heated at 2° C./minfrom −80° C. to 300° C. A purge of dry nitrogen at 50 mL/min wasmaintained over the sample. Modulated temperature DSC was carried outusing an underlying heating rate of 2° C./min and temperature modulationparameters of ±1.272° C. (amplitude) every 60 seconds (period). Theinstrument control software was Advantage for Q Series v2.8.0.392 andThermal Advantage v4.8.3 and the data were analyzed using UniversalAnalysis v4.4A.

Non-modulated DSC data were collected on a TA Instruments Q2000 equippedwith a 50 position auto-sampler. The calibration for thermal capacitywas carried out using sapphire and the calibration for energy andtemperature was carried out using certified indium. Typically 1 to 5 mgof each sample, in an aluminum pan, was heated at 10° C./min from 20 OCto 300° C. A purge of dry nitrogen at 50 mL/min was maintained over thesample. The instrument control software was Advantage for Q Seriesv2.8.0.392 and Thermal Advantage v4.8.3 and the data were analyzed usingUniversal Analysis v4.4A.

6.1.1.3 Thermogravimetric Analysis (TGA)

TGA data were collected on a Mettler TGA/SDTA 851e equipped with a 34position autosampler. The instrument was temperature calibrated usingcertified indium. Typically 5-15 mg of each sample was loaded onto apre-weighed aluminum crucible and was heated at 10° C./min from ambienttemperature to 350° C. A nitrogen purge at 50 ml/min was maintained overthe sample. The instrument control and data analysis software was STARev9.20.

6.1.1.4 Polar Light Microscopy

Samples were studied on a Leica LM/DM polarized light microscope with adigital video camera for image capture. A small amount of each samplewas placed on a glass slide, mounted in immersion oil and covered with aglass slip, the individual particles being separated as well aspossible. The sample was viewed with appropriate magnification andpartially polarised light, coupled to a λ false-colour filter.

6.1.1.5 Gravimetric Vapour Sorption (GVS)

Sorption isotherms were obtained using a SMS DVS Intrinsic moisturesorption analyser, controlled by DVS Intrinsic Control softwarev1.0.0.30. The sample temperature was maintained at 25° C. by theinstrument controls. The humidity was controlled by mixing streams ofdry and wet nitrogen, with a total flow rate of 200 mL/min. The relativehumidity was measured by a calibrated Rotronic probe (dynamic range of1.0-100% RH), located near the sample. The weight change, (massrelaxation) of the sample as a function of % RH was constantly monitoredby the microbalance (accuracy±0.005 mg). Typically 5-20 mg of sample wasplaced in a tared mesh stainless steel basket under ambient conditions.The sample was loaded and unloaded at 40% RH and 25° C. (typical roomconditions). The standard isotherm was performed at 25° C. at 10% RHintervals over a 0-90% RH range. Data analysis was undertaken inMicrosoft Excel using DVS Analysis Suite v6.0.0.7.

6.1.2 Solid Form Screen Experiments

The solvents used in the polymorph screen were either HPLC or reagentgrade, including toluene, MTBE (methyl tert-butyl ether), DIPE(diisopropyl ether), THF (tetrahydrofuran), DME (dimethoxyethane), IPAc(isopropyl acetate), EtOAc (ethyl acetate), MIBK (methyl isobutylketone), acetone, IPA (isopropyl alcohol), ethanol, ACN (acetonitrile),nitromethane, p-xylene, p-xylene/acetone (e.g., 50:50), p-xylene/MTBE(e.g., 50:50), or IPA:water (e.g., 95:5).

The solid form generated from the screen was characterized by X-raypowder diffraction (XRPD), differential scanning calorimetry (DSC),thermogravimetric analysis (TGA), optical microscopy, and gravimetricvapor sorption (GVS).

6.1.2.1 Equilibration/Slurry and Evaporation

Amorphous Compound A (˜10 mg per experiment) was treated with the statedsolvent. Solutions were allowed to slowly evaporate at room temperatureand the residual solids were analyzed by XRPD. Suspensions weresubjected to heat/cool cycles (50° C./room temperature, 8 hour cycle)for 16 hours; the solvent was then allowed to evaporate and the residualsolids were analyzed by XRPD.

The results of slurry experiments are summarized in Table 1. All of thesolids obtained from filtration of the slurries were confirmed to beForm A by XRPD.

TABLE 1 Slurry Experiments of Form A of Compound A at Room TemperatureSolvent XRPD Result Toluene Form A MTBE Form A DIPE Form A THF Form ADME Form A IPAc Form A EtOAc Form A MIBK Form A Acetone Form A IPA FormA Ethanol Form A ACN Form A Nitromethane Form A IPA:water (95:5) Form A

6.1.3 Characterization of Form A of Compound A

6.1.3.1 XRPD, TGA, and DSC Characterization

Form A has a crystalline XRPD pattern as shown in FIG. 1 and anirregular plate crystal habit as shown in FIG. 2. The XRPD pattern ofForm A of Compound A shows that Form A is crystalline. Some XRPD peaksof crystalline Form A are summarized in Table 2.

TABLE 2 X-Ray Diffraction Peaks for Form A of Compound A Two-theta angle(°) d Space (Å) Intensity (%) 8.3 10.648 58.3 8.8 9.984 26.8 12.0 7.3428.1 13.2 6.708 100.0 13.9 6.357 8.0 14.4 6.125 3.3 14.8 5.961 8.7 16.55.352 50.2 17.7 4.996 35.4 18.2 4.872 50.7 19.3 4.586 8.2 19.5 4.560 7.719.6 4.526 7.3 21.0 4.230 4.4 21.2 4.185 3.9 21.7 4.094 50.9 22.5 3.94213.6 24.1 3.684 8.4 24.7 3.603 7.1 25.0 3.560 12.8 25.3 3.512 5.6 26.53.363 35.7 26.7 3.332 5.7 28.3 3.147 11.4 29.3 3.051 5.5 29.5 3.022 9.929.8 2.992 7.9 30.5 2.924 3.2 32.1 2.782 2.9 33.3 2.690 3.4 34.2 2.6213.5 34.6 2.587 4.4

TGA and DSC thermograms of Form A are shown in FIG. 3. Form A was foundto lose up to 0.02% volatiles during TGA analysis upon 100° C., whichindicates that Form A is unsolvated and anhydrous. Form A exhibited asingle melting peak at 199.3° C. (onset).

6.1.3.2 Hygroscopicity

Hygroscopicity of Form A was determined by moisture adsorption anddesorption. The moisture sorption/desorption behavior of Form A wasdetermined by DVS and the results are summarized in FIG. 4. Form Ashowed no significant water uptake (<0.1% w/w) between 0 and 80%relative humidity, which indicates that Form A is not hygroscopic. Afterundergoing the full adsorption/desorption cycle, the XRPD diffractogramof the sample showed that the material was unchanged from the initialForm A. Based on the characterization results, Form A was found to be ananhydrous and non-hygroscopic crystalline material.

6.1.4 Alternative Methods for the Preparation of Form A of Compound A

Preparation 1:

Compound A was combined with BHT (0.001 equiv) in IPA and water (3×:5×vol). The mixture was heated 65° C. and while maintaining thistemperature, water (5× vol) heated to 65° C. was added. A small amountof the title compound (0.02 equiv) in water heated to 65° C. was added.The mixture was held for 2 h, cooled to room temperature over 4 h, andstirred for an additional 2 h. The resulting solids were collected byfiltration, washed with 20% IPA in water and dried to give Compound A asa white to yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm) 9.03 (d,J=1.56 Hz, 1H), 8.28 (s, 1H), 8.24 (dd, J=2.34, 8.20 Hz, 1H), 7.74 (d,J=7.81 Hz, 1H), 7.61 (s, 1H), 5.26 (s, 1H), 4.90 (tt, J=3.71, 12.10 Hz,1H), 4.13 (s, 2H), 3.28 (s, 3H), 3.20 (tt, J=4.00, 10.84 Hz, 1H), 2.58(qd, J=2.93, 12.82 Hz, 2H), 2.14 (d, J=10.15 Hz, 2H), 1.68 (d, J=10.93Hz, 2H), 1.47 (s, 6H), 1.17-1.35 (m, 2H); MS (ESI) m/z 398.3 [M+1]⁺. DSCendotherm at 201.9° C. XRPD diffractogram (top peaks±0.5°) two-thetaangle (°): 8.0, 9.0, 12.0, 13.0, 16.5, 17.5, 18.2, 21.5, 22.5, 25.0,26.5.

Preparation 2:

Compound A was combined with BHT (0.02 equiv) in MeOAc (25× vol) andheated to 55° C. The solution was cooled to 25° C. and a small amount ofthe title compound (0.02 equiv) in MeOAc was added. The slurry was heldfor 1 h, distilled under vacuum to a reduced volume and treated withn-heptane (10× vol). The slurry was held for 2 h, and the resultingsolids were collected by filtration, washed with 50% MeOAc in n-heptaneand dried to give Compound A as a white to yellow solid. ¹H NMR (400MHz, DMSO-d₆) δ (ppm) 9.03 (d, J=1.56 Hz, 1H), 8.28 (s, 1H), 8.24 (dd,J=2.34, 8.20 Hz, 1H), 7.74 (d, J=7.81 Hz, 1H), 7.61 (s, 1H), 5.26 (s,1H), 4.90 (tt, J=3.71, 12.10 Hz, 1H), 4.13 (s, 2H), 3.28 (s, 3H), 3.20(tt, J=4.00, 10.84 Hz, 1H), 2.58 (qd, J=2.93, 12.82 Hz, 2H), 2.14 (d,J=10.15 Hz, 2H), 1.68 (d, J=10.93 Hz, 2H), 1.47 (s, 6H), 1.17-1.35 (m,2H); MS (ESI) m/z 398.3 [M+1]⁺. DSC endotherm at 201.9° C. XRPDdiffractogram (top peaks, +0.5°) two-theta angle (°): 8.0, 9.0, 12.0,13.0, 16.5, 17.5, 18.2, 21.5, 22.5, 25.0, 26.5

Preparation 3:

Compound A was combined with BHT (0.02 equiv), and MeOAc, and heated to55° C., forming a clear solution. The solution was filtered while hot,cooled to 30° C. and a small amount of the title compound (0.02 equiv)in MeOAc was added. The slurry was agitated for at least 1 h, distilledunder vacuum to a reduced volume and treated with n-heptane. Theresulting solid was collected through filtration, washed with a 1:1mixture of MeOAc in n-heptane and dried to give Compound A as a white toyellow solid ¹H NMR (400 MHz, DMSO-d₆) δ (ppm) 9.03 (d, J=1.56 Hz, 1H),8.28 (s, 1H), 8.24 (dd, J=2.34, 8.20 Hz, 1H), 7.74 (d, J=7.81 Hz, 1H),7.61 (s, 1H), 5.26 (s, 1H), 4.90 (tt, J=3.71, 12.10 Hz, 1H), 4.13 (s,2H), 3.28 (s, 3H), 3.20 (tt, J=4.00, 10.84 Hz, 1H), 2.58 (qd, J=2.93,12.82 Hz, 2H), 2.14 (d, J=10.15 Hz, 2H), 1.68 (d, J=10.93 Hz, 2H), 1.47(s, 6H), 1.17-1.35 (m, 2H); MS (ESI) m/z 398.3 [M+1]⁺. DSC endotherm at201.9° C. XRPD diffractogram (top peaks, ±0.5°) two-theta angle (°):8.0, 9.0, 12.0, 13.0, 16.5, 17.5, 18.2, 21.5, 22.5, 25.0, 26.5.

Preparation 4:

A 1:1 wt/wt mixture of Compound A (Form A) and Compound A (pinacolco-crystal) was treated with IPA (6× vol) with agitation for 4 days atambient temperature. The solids were collected by filtration and driedunder reduced pressure at 40-50° C. to give Compound A (Form A) as ayellow solid. DSC endotherm of 195° C. XRPD diffractogram (top peaks,+0.5°) two-theta angle (°): 8.0, 9.0, 12.0, 13.0, 16.5, 17.5, 18.2,21.5, 22.5, 25.0, 26.5.

Preparation 5:

Compound A was combined with BHT (0.02 equiv) and 5% aqueous THF (5×vol) to form a clear solution. The solution was optionally treated withactivated carbon for 4 h. The solution was filtered, treated withisopropyl acetate (3× vol), and distilled at atmospheric pressure atconstant volume with addition of isopropyl acetate until the solutiontemperature reached 80° C. The solution was cooled to 75° C. and treatedwith a small amount of the title compound (0.02 equiv) in isopropylacetate, and the slurry was held for 2 h. The slurry was distilled atatmospheric pressure at constant volume with addition of isopropylacetate until the slurry temperature reached 88° C. The slurry wascooled to 80-85° C., held for 2 h, cooled to 25° C. over 4 h, and heldfor at least 8 h. The resulting solid was collected through filtration,washed with isopropyl acetate, and dried to give Form A as a white toyellow solid. Alternatively, after filtration and addition of isopropylacetate, the distillation at constant volume can be carried out underreduced pressure keeping the reaction mixture at 40° C. until isopropylacetate (3.5× vol) has been added. The solution at 40° C. is thentreated with a small amount of the title compound (0.02 equiv) inisopropyl acetate, held for 2 h, and distilled at constant volume undervacuum at 40° C. with addition of an additional portion of isopropylacetate (10× vol). The slurry is aged at 40° C. for 2 h, cooled to 25°C. over 2 h, held for at least 8 h, and the solid collected throughfiltration, washed with isopropyl acetate, and dried to give Form A as awhite to yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm) 9.03 (d, J=1.56Hz, 1H), 8.28 (s, 1H), 8.24 (dd, J=2.34, 8.20 Hz, 1H), 7.74 (d, J=7.81Hz, 1H), 7.61 (s, 1H), 5.26 (s, 1H), 4.90 (tt, J=3.71, 12.10 Hz, 1H),4.13 (s, 2H), 3.28 (s, 3H), 3.20 (tt, J=4.00, 10.84 Hz, 1H), 2.58 (qd,J=2.93, 12.82 Hz, 2H), 2.14 (d, J=10.15 Hz, 2H), 1.68 (d, J=10.93 Hz,2H), 1.47 (s, 6H), 1.17-1.35 (m, 2H); MS (ESI) m/z 398.3 [M+1]⁺. DSCendotherm at 201.9° C. XRPD diffractogram (top peaks, +0.5°) two-thetaangle (°): 8.0, 9.0, 12.0, 13.0, 16.5, 17.5, 18.2, 21.5, 22.5, 25.0,26.5.

6.1.5 Preparation of Pinacol Co-Crystal of Compound A

Compound A, pinacol (2.4 equiv), and THF (5× vol) were combined andheated to 45-50° C., and toluene (lx vol) was added. The solution wasdistilled under reduced pressure (300-350 Torr) keeping the temperaturebetween 40-45° C. to 4× vol. The solution was cooled, and toluene (5×vol) was added while continuously removing solvent under reducedpressure (300-350 Torr), until 15% THF in toluene composition wasachieved. The batch was seeded with pinacol co-crystal (0.02 equiv) at25° C., and the batch was held for 72 h. The solids were filtered,rinsed with THF/toluene and dried at 45-50° C. under vacuum to affordCompound A pinacol co-crystal (71% yield, 20 wt % pinacol by ¹H NMR).DSC melt at 119.0° C. XRPD diffractogram (top peaks, ±0.5°) two-thetaangle (°): 5.0, 6.0, 12.5, 14.0, 15.0, 15.5, 17.5, 18.5, 22.5.

6.1.6 Preparation of Hydrate of Compound A (Form B)

Compound A was combined with BHT (0.001 equiv) in IPA and water (3×:5×vol). The mixture was heated to 55° C., and water (5× vol) was added. Asmall amount of the title compound (0.02 equiv) in water was added. Themixture was cooled to room temperature over 1 h and stirred for anadditional 48 h at room temperature. The resulting solids were collectedby filtration, washed with 20% IPA in water and dried to give Compound Ahydrate as a pink solid. The solid had a DSC endotherm of 111.3° C.,exotherm of 164.9° C., and endotherm of 201.6° C. TGA analysis showed6.4% weight loss and an onset temperature of 50° C. XRPD diffractogram(top peaks, +0.5°) two-theta angle (°): 6.0, 7.0, 8.0, 10.0, 12.0, 14.0,17.0, 18.0, 20.0, 20.5, 22.5, 24.5.

6.1.7 Preparation of Anhydrous Form of Compound A (Form C)

Preparation 1:

Compound A was combined with BHT (0.001 equiv) in MeOH (10× vol). Themixture was distilled to a reduced volume (5×) and further distilledwith the addition of IPA until an additional 50 mL of distillate wascollected, and the solution was cooled to room temperature. Theresulting solids were collected by filtration, washed with IPA, (2× vol)and dried to give Compound A as an off-white solid. DSC analysis of thesolid showed an endotherm of 161° C. and an endotherm of 200° C. XRPDdiffractogram (top peaks, +0.5 0) two-theta angle (°): 6.5, 9.0, 10.0,14.5, 16.5, 19.0, 23.0, 23.5.

Preparation 2:

Compound A (pinacol co-crystal) and BHT (0.01× wt) were treated with IPA(8× vol) with agitation for 4 days at ambient temperature. The solidswere collected by filtration, washed with IPA, and dried under reducedpressure at 40-50° C. to give Compound A (Form C) as a solid. DSCanalysis of the solid showed an endotherm and exotherm at 160° C. and anendotherm at 200° C. XRPD diffractogram (top peaks, ±0.5°) two-thetaangle (°): 6.5, 9.0, 10.0, 14.5, 16.5, 19.0, 23.0, 23.5.

6.1.8 Preparation of Methanol Solvate of Compound A (Form D)

Compound A was combined with BHT (0.001 equiv) in MeOH (20× vol) andheated to 65° C. The solution was cooled to room temperature and stirredfor an additional 18 h. The resulting solids were collected byfiltration, washed and dried at 40-45° C. to give Compound A as a pinksolid. The solid had a DSC endotherm of 98.3° C., an exotherm of 159.3°C., and an endotherm of 200.6° C. TGA analysis showed 7.4% weight lossand an onset temperature of 80° C. XRPD diffractogram (top peaks, ±0.5°)two-theta angle (°): 6.0, 7.5, 8.0, 9.0, 10.0, 12.5, 14.5, 16.5, 19.0,19.5, 20.5, 23.0.

6.1.9 Preparation of p-Xylene Solvate of Compound A (Form E)

The XRPD pattern, crystal habit, TGA, SDTA, TGA-MS, HPLC and MS of FormE of Compound A are shown in FIGS. 15-20. Form E was prepared in atleast three experiments, including: a slurry conversion experiment inp-xylene, a hot-filtration experiment in acetone/p-xylene (50/50), andan evaporative experiment in MTBE/p-xylene (50/50). The samples used forfurther analyses were prepared in the slurry conversion experiment inp-xylene.

FIG. 15 provides an overlay of XRPD patterns (from bottom to top) of:starting material (Form A of Compound A), Form E as a wet solid obtainedfrom a slurry conversion experiment in p-xylene, Form E as a driedsolid, Form E as a wet solid obtained from a slurry conversionexperiment in p-xylene after exposure to accelerated aging conditions(AAC) and Form E as a dried solid after exposure to AAC. A list of X-RayDiffraction Peaks for Form E of Compound A is provided below in Table 3.

TABLE 3 X-Ray Diffraction Peaks for Form E of Compound A Two-theta angle(°) d Space (Å) Relative Intensity (%) 7.46 11.84 86.65 8.94 9.88 20.9211.7 7.55 13.33 13.7 6.46 24.65 17.26 5.13 14.44 18.22 4.86 21.32 18.784.72 20.1 20.94 4.24 11.48 22.38 3.97 19.83 23.06 3.85 23.69 24.62 3.6118.4

A single-crystal X-ray diffraction analysis is employed to determine thecrystal structure of Form E of Compound A. Table 4 and Table 5 present asummary of the crystallographic data from the crystal-structuredetermination. Form E of Compound A has a crystal packing pattern asshown in FIG. 16. From the crystal structure results, it is confirmedthat Form E is a p-xylene hemi-solvated form crystallizing in amonoclinic crystal system.

TABLE 4 Crystal data and structure refinement of Form E Empiricalformula C₂₁H₂₇N₅O₃•0.5 C₈H₁₀ Fw 450.56 T [K] 296(2) λ [Å] 0.71073Crystal system Monoclinic Space group P 2₁/c Unit cell dimensions a [Å]12.3583(6) b [Å] 15.1360(9) c [Å] 13.4604(6) β [°] 105.272(4) V [Å³]2428.9(2) Z 4 D_(c) [g/cm³] 1.232 μ [mm⁻¹] 0.083 F(000) 964 Crystal size[mm³] 0.35 × 0.26 × 0.12 θ range for data collection [°] 2.5 → 32.6Reflections collected 26665 Independent reflections 8818 [R_(int) is0.0361] Completeness to θ = 32.6° [%] 99.3 Max. and min. transmission0.9901 and 0.9716 Data/restraints/parameters 8818/0/406 Goodness-of-fiton F² 1.043 Final R indices [I > 2σ(I)] R1 = 0.0531, wR2 = 0.1342 Rindices (all data) R1 = 0.0785, wR2 = 0.1528

TABLE 5 Hydrogen bonds of Form E D-H . . . A D-H H . . . A D . . . A D-H. . . A [Å] [Å] [Å] [°] O(1)—H(1) . . . N(13)^(i) 0.89(2) 2.01(2)2.865(2) 162(2) N(15)—H(15) . . . N(6)^(ii) 0.86(2) 2.14(2) 2.976(2)164(2)

FIG. 17A is a digital image of Form E as a wet solid obtained fromslurry conversion experiment in p-xylene. FIG. 17B is a digital image ofForm E as a dried solid. FIG. 17C is a digital image of Form E as a wetsolid obtained from slurry conversion experiment in p-xylene afterexposure to AAC. FIG. 17D is a digital image of Form E as a dried solidafter exposure to AAC.

FIG. 18 and FIG. 19 provide TGA/SDTA signal and TGA-MS data,respectively, of Form E.

The TGA thermogram of Form E in FIG. 18 shows a mass loss correspondingto a broad endothermic event observed in the SDTA signal between 90° C.and 125° C. with a maximum at about 106-110° C., which may be thedesolvation of Form E. After the desolvation, the SDTA data of Form E inFIG. 18 shows a melting event at 193° C., corresponding to the meltingof the starting material, Form A of Compound A.

The TGA thermogram of Form E in FIG. 19 comprises a total mass loss ofapproximately 11.0% of the total mass of the sample betweenapproximately 50° C. and approximately 180° C. when heated fromapproximately 25° C. to approximately 300° C. Thus, Form E loses about11.0% of its total mass when heated from about ambient temperature toabout 300° C. The thermal data indicates that Form E contains 0.5 molarequivalents of solvent in the crystal lattice corresponding toapproximately 0.5 mole of p-xylene per mole of Compound A. Thetheoretical p-xylene content of a p-xylene hemi-solvate of Compound A is10.5% by weight, matching the TGA weight loss observed. Theseobservations suggest that Form E is a p-xylene hemi-solvate of CompoundA.

FIG. 20 provides HPLC and MS data of Form E. The peak retention time is8.1 minutes with a sample purity of 98.9% (area %).

6.2 Synthesis

6.2.1 Large Scale Synthesis of Compound A

6.2.1.1 Synthesis 1

Ethyl-2-(3,5-dibromopyrazin-2-ylamino)acetate (70.0 kg),trans-4-methoxycyclohexylamine hydrochloride, (51.5 kg) and NMP (360.1kg) were combined and treated with DIPEA (93.5 kg). The batch was heatedto 125-130° C. until completion was reached. The resulting reactionmixture was cooled to 20-35° C. and quenched into a mixture of 5% sodiumchloride solution and EtOAc. The organic layer was washed three timeswith a 5% sodium chloride solution followed by a water wash. The organicphase was concentrated by distillation, causing the solid product toform. The solid was collected through filtration, washed with MTBE anddried (40% yield).

Ethyl2-((5-bromo-3-(((trans)-4-methoxycyclohexyl)amino)pyrazin-2-yl)amino)acetate(35.0 kg) was treated with a 21% phosphoric acid solution (147.4 kg) at80° C. for at least 12 h. The resulting suspension was cooled to roomtemperature an d the solid was collected through filtration and washedwith water. The solid was slurried in water and treated with a 1 Mpotassium carbonate solution (1 equiv, 12.6 kg). The resulting solid wascollected through filtration, washed with water, and dried (85% yield).

7-Bromo-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one(27.5 kg),2-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)propan-2-olhydrochloride (26.2 kg), and PdCl₂(Amphos)₂ (137.5 g) in THF (219.8 kg)were combined with a potassium carbonate solution (27.5 kg), and heatedto reflux until reaction completion was reached. The mixture was cooled,treated with toluene, and the aqueous layer was removed. The organicsolution was washed with an aqueous potassium dihydrogen phosphatesolution, and the aqueous layer was removed. The organic layer wastreated with SiliaBond® Thiol (4.2 kg) and twice with activated carbon(2×2.8 kg). The organic solution was distilled to a reduced volumefollowed by continuous distillation with the addition of toluene until a15% THF in toluene solution was reached, at which time the batch wascooled and the product was left to precipitate. The resulting solid wascollected through filtration, washed with toluene, and dried (70%yield).

6.2.1.2 Synthesis 2

A mixture of ethyl-2-(3,5-dibromopyrazin-2-ylamino)acetate (69.1 kg),trans-4-methoxycyclohexylamine hydrochloride, (50.8 kg) and NMP (360 kg)was heated to 125-130° C. until completion was achieved. The mixture wascooled to 20-30° C., and treated with 5% sodium chloride solution (5vol) and EtOAc (8 vol). The aqueous layer was removed, and the organiclayer was washed three times with 5% sodium chloride (3×5 vol) and oncewith water (5 vol). The organic layer was concentrated by vacuumdistillation to a reduced volume, cooled to 25° C., and agitated at thistemperature for 19 h. The slurry was filtered and the wet cake waswashed with MTBE. The product was dried in a vacuum oven at to obtainethyl2-((5-bromo-3-(((trans)-4-methoxycyclohexyl)amino)pyrazin-2-yl)amino)acetate(44% yield). Alternatively, the reaction can be carried out using NMP(3× vol) and after workup and distillation to a reduced volume, thereaction mixture can be held at 50° C., seeded with a small amount ofthe target compound (0.02 equiv), treated with n-heptane (3× vol),cooled to 25° C. over 2 h, and aged at least 12 h. The solids can becollected by filtration, washing with a EtOAc/n-heptane mixture, anddrying (75% yield).

Ethyl2-((5-bromo-3-(((trans)-4-methoxycyclohexyl)amino)pyrazin-2-yl)amino)acetate(35 kg) was treated with a 21% phosphoric acid solution (410 kg) at 80°C. until completion was achieved. The suspension was cooled to 30-35° C.and filtered, and the wet cake was washed with water (5× vol), chargedto a reactor, and suspended in water (3× vol). The slurry was treatedwith 1M potassium carbonate solution (1 equiv), filtered and washed withwater (2×5× vol). The product was dried at 50-55° C. in a vacuum oven todeliver7-bromo-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one(91% yield).

A mixture of7-bromo-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one(also having the chemical name7-bromo-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one)(27.7 kg),2-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)propan-2-olhydrochloride (26.3 kg) and PdCl₂(Amphos)₂ (137.6 g) in THF (122.7 kg)was combined with a solution of potassium carbonate (27.5 kg) in water(220 kg). The mixture was heated to reflux and held until reactioncompletion. The batch was cooled to 45° C., toluene (71.4 kg) was added,and the aqueous phase was removed. The organic solution was treated withaqueous potassium dihydrogen phosphate solution, SiliaBond® Thiol, andtwice with activated carbon. The resulting organic solution wasdistilled under atmospheric pressure to a reduced volume andcontinuously distilled with toluene addition until a composition of ˜15wt % THF in toluene was reached. The batch was cooled to 25° C.,filtered, and the solids were washed with toluene, and dried undervacuum to deliver Compound A as a light yellow solid (87% yield).

Alternatively, after reaction completion the batch can be treated withtert-butyl ether (MTBE), washed with aqueous potassium dihydrogenphosphate solution, treated with SiliaBond Thiol, and activated carbon,followed by distillative solvent exchange to isopropanol under eitheratmospheric or reduced pressure to afford crystallization, followed bythe addition of n-heptane, filtration and drying under vacuum to affordCompound A (90% yield). Without being limited by theory, it is thoughtthat the distillative solvent exchange to isopropanol can avoid orreduce formation of a pinacol co-crystal, thus providing Compound A ofhigher purity.

Compound A (27.1 kg), BHT, (270 g) and MeOAc (604 kg) were combined,heated to 50-55° C., and filtered. A slurry of small amount of CompoundA (540 g) in MeOAc (2.6 kg) was added, and the batch was held for 1 h.The batch was distilled under vacuum to 10× vol, and treated withheptane while maintaining the batch temperature at 25-30° C. until thecomposition is 1:1 (v/v/) MeOAc/heptane. The batch was held at 20-25° C.for 14 h, filtered, and the wet cake was washed twice with 1:1MeOAc/heptane and dried at 50-55° C. under vacuum to deliver Compound A(78% yield) as an off-white to light yellow solid. DSC confirmed thecrystal Form A. ¹H NMR (DMSO-d₆) was consistent with the assignedstructure.

6.2.2 Large Scale Synthesis of Metabolite of Compound A

A metabolite of Compound A was prepared as follows:

A vessel was charged with 1 (2.15 kg), 2 (1.44 kg), and NMP (6.5 L), andthe resulting slurry was agitated at 20-30° C. and treated with DIPEA(3.87 L). The batch was heated to 125-130° C., held for 20 hours untilcompletion was achieved, cooled to 20-35° C., and transferred to avessel containing a mixture of EtOAc (17.2 L) and 5% aq. NaCl (10.7 L).The batch was agitated for 10-15 minutes, allowed to settle for 10-15minutes, and the aqueous layer was removed. The batch was washed anadditional three times with 5% aq. NaCl (10.7 L) and once with water(10.7 L). The batch was distilled under reduced pressure (50-60° C.;250-300 Torr) until reaching 2× volume. The resulting slurry was treatedwith n-heptane (6.3 L) while maintaining a batch temperature of 50-60°C. The batch was cooled to 20-30° C., held for 17 hours, and filtered.The filter cake was washed with n-heptane and dried at 50-60° C. undervacuum to afford 3 (66% yield) as a solid.

The solid 3 (1.56 kg) and a 10% aq. H₃PO₄ solution (16 L) were heated to75-85° C., held for 15 hours, cooled to 20-30° C., and filtered. Thefilter cake was washed with water (5 L) and dried on the filter for 1hour. The filter cake was charged to a vessel, treated with water (15L), and agitated at 20-30° C. for 2 hours. The batch was filtered,washed with water (2×4.7 L), dried in a vacuum oven at 50-60° C. toobtain 4 (54% for two steps) as solid. MS: Calc: 327.0 [M+H]; Obsd:309.0 [M-OH], 329.0 [M+3].

A vessel was charged with 4 (447 g), 5 (425 g), PdAmphos₂Cl₂ (0.00023eq.), and THF (2.2 L) that had been sparged with N₂ for 30 min. Theslurry was agitated and treated with a solution of K₂CO₃ (2.4 eq.) inwater (3.6 L), that had been sparged with N₂ for 30 min. The batch washeated to reflux, held for 15 h, cooled to just below the reflux point,and an additional charge of PdAmphos₂Cl₂ (0.00046 eq.) was added. Themixture was heated to reflux, held for 20 h, cooled to 40-50° C.,treated with toluene (447 mL), and the aqueous layer was removed. Thebatch was treated with toluene (447 mL) at which time precipitation ofsolids began. The batch was distilled under atmospheric pressure to 6×vol and distilled at constant volume with addition of toluene until thecomposition reached ˜30% THF in toluene.

The supernatant was removed, and the remaining solids were treated withTHF (447 mL), heated to 60-65° C., and treated with THF (447 mL). Thebatch was held at 60-65° C. for 30 minutes, cooled to 20-30° C. over 45minutes, and aged for 15 hours at 20-30° C. The batch was treated withTHF (447 mL) and filtered. The filter cake was dried under vacuum at40-50° C. to obtain crude 6 (59% yield) as a solid. MS: Calcd: 384.2[M+H]; Obsd: 384.2.

The THF filtrate was concentrated under reduced pressure, slurried inIPA (500 mL) for 4 hours and filtered. The filtered solids were driedunder vacuum at 40-50° C. to obtain crude 6 (23% yield) as a solid. MS:Calcd: 384.2 [M+H]; Obsd: 384.2.

A vessel was charged with crude 6 (310 g), BHT (155 mg), SiliaBond®Thiol (47 g), THF (11.8 L), and water (620 mL) and agitated to form aslurry. The batch was heated to 50-55° C., held for 4 hours, cooled to30-40° C., and filtered. The filtrate was charged to a vessel distilledunder reduced pressure (27-30° C., 200 mmHg) until reaching 5-6× vol.The batch was cooled to 20-30° C., agitated for 2 hours, and filtered.The filter cake was washed with THF (300 mL) and dried under vacuum at45-50° C. The resulting solid (153 g), BHT (75 mg), IPA (1.1 L), andwater (380 mL), were combined and agitated to form a slurry. The slurrywas heated at elevated temperature (reflux) for 18 h, cooled to 20-30°C., held for 3-4 hours, and filtered. The filter cake was dried at 50°C. under vacuum to deliver purified 6 (66% yield) as a solid. MS: Calcd:384.2 [M+H]; Obsd: 384.2.

6.3 Synthesis of Isotopologues of Compound A

6.3.1 Synthesis of ¹⁴C Enriched Compound A

¹⁴C-radiolabeled Compound A was prepared as follows.

5-Bromo-2-iodopyridine (1 equiv) in DCM was cooled to −78° C. andtreated sequentially with n-BuLi (1.05 equiv of 2.5M in hexane) and¹⁴C-labeled acetone (3 equiv). The mixture was slowly warmed to ambienttemperature, stirred for 30 min, and treated with water (10 mL). Theorganic layer was dried with Na₂SO₄, filtered, and concentrated underreduced pressure. The crude product was taken to the next step with noadditional purification.

Crude 7 in DCM at ambient temperature was sequentially treated with TEA(3 equiv) and TMSCl (2 equiv) and stirred for 18 h. The reaction mixturewas treated with saturated NaHCO₃ (15 mL), and extracted with DCM. Theorganic layer was dried with Na₂SO₄, filtered, and concentrated underreduce pressure. The oil was purified by column chromatography (5%EtOAc/hexane) to deliver 8 as an oil (52% over 2 steps).

Compound 8, bis(pinacolato)diborane (1.1 equiv), KOAc (3 equiv), andPdCl₂(dppf)-DCM complex (0.03 equiv) were combined in 1,4-dioxane,heated to 90° C., and held for ˜18 h. The mixture was cooled to ambienttemperature, diluted with MTBE, filtered, and concentrated under reducedpressure. The crude material was purified by column chromatography (1:1EtOAc:hexane) to obtain Compound 9 as a solid (27% yield).

Compound 9 in 1,4-dioxane was treated with 4 M HCl in 1,4-dioxane (2equiv) at ambient temperature and stirred for 2 h. The mixture wasconcentrated under a flow of N₂ to give an off white solid, which wastreated with MTBE for 1 h and filtered to obtain Compound 10 as a solid(98% yield).

Compound 10, Compound 11 (1.08 equiv), PdCl₂(Amphos)₂ (0.02 equiv), THF,and an aqueous K₂CO₃ solution (2.5 equiv K₂CO₃) were heated in a sealedtube at 70-75° C. for 16 h. The tube was cooled to 25° C., and themixture was extracted with toluene and concentrated under reducedpressure. The crude oil was purified by column chromatography (1:1THF/DCM) and isocratic semi-preparative HPLC. The isolated fractionswere concentrated under reduced pressure, dissolved in EtOAc, dried withNa₂SO₄, filtered, and concentrated under reduced pressure. The materialwas dissolved in THF and concentrated under a flow of nitrogen followedby high vacuum. The isolated oil was treated with ACN and concentratedwith a stream of N₂ to induce crystallization. The contents of wereconcentrated under high vacuum to obtain ¹⁴C-labeled Compound A as asolid.

Alternatively, ¹⁴C-Compound A can be prepared from 10 and 11 as follows:

Compound 10 and 11 (1.1 equiv), THF, and aqueous K₂CO₃ (2.5 equivK₂CO₃), were combined with PdAmphos₂Cl₂ (0.02 equiv) and heated to70-75° C. until reaction completion (about 18 h). The mixture wascooled, treated with EtOAc and brine and the layers separated. Theorganic layer was dried over Na₂SO₄, filtered, and concentrated to aresidue. The residue was purified by column chromatography on silica gel(CH₂Cl₂:EtOAc 1:3; followed by MeOH:EtOAc 2:98) and concentrated to aresidue. The residue was then purified by preparative HPLC using 0.015 MKH₂PO₄ and MeCN. The collected fractions were extracted with EtOAc,dried over Na₂SO₄, filtered, and concentrated to obtain ¹⁴C-labeledCompound A as a solid.

6.3.2 Synthesis of ¹³C Enriched Compound A

¹³C-labeled Compound A was prepared as follows.

K₂CO₃ (1.5 eq,) and ethyl bromoacetate-¹³C₂ (1.3 eq) were added to asolution of 3,5-dibromopyrazin-2-amine (1.0 eq) in acetone (10× vol).The slurry was heated to 30° C., Bu₄NHSO₄ (0.074 eq) was added, and themixture was stirred for 2 d at reflux. The reaction slurry was cooled toambient temperature, filtered through celite, and the cake was washedwith acetone (10 vol). The filtrate was concentrated under reducedpressure, dissolved in EtOAc (11.4 vol), and the organic phase waswashed with water (2×3.2 vol) and saturated aqueous NaCl (2×3.2 vol).The combined aqueous phase was extracted with EtOAc, and the combinedorganic phase was dried over MgSO₄, filtered, and washed with EtOAc.Ecosorb-906 (0.11 wt) was added, and the mixture was stirred 13 h. Theslurry was filtered washed with EtOAc, and the filtrate was concentratedunder reduced pressure to a slurry to which was added a 2% EtOAc inheptane solution (7.9 vol). The slurry was filtered after stirring for 3h at ambient temperature. The collected solid was washed with heptane (3vol) and dried in a vacuum oven at 35° C. to provide (12) as a solid(57% yield). ¹H NMR (CDCl₃, 300 MHz): δ=8.05 (s, 1H), 5.77 (br. s., 1H),4.41 (t, J=5.7 Hz, 1H), 4.26 (qd, J=7.1, 3.0 Hz, 2H), 3.94 (t, 1H), 1.31(t, J=7.1 Hz, 3H) ppm. LC/MS: Calculated: 340.9, Found: ES+ (M+1) 341.9.

A reaction flask was sequentially charged withtrans-4-methoxycyclohexanamine hydrochloride (1.5 eq), compound (12)(1.0 eq), NMP (5.0 vol) and DIPEA (3.5 eq). The solution was heated to125° C. for 24 h and then cooled to 25° C. EtOAc (10 vol) and 5% aqueousNaCl (15 vol) were added, and the layers were separated. The organiclayer was washed with a 5% aqueous NaCl (2×15 vol) and concentratedunder reduced pressure. The residue was treated with MTBE (4.0 vol),stirred 1 hour at ambient temperature and filtered. The solid was washedwith MTBE and dried in a vacuum oven at 20-30° C. to provide (13) as asolid (61% yield). ¹H NMR (DMSO-d₆, 300 MHz): δ=7.21 (s, 1H), 6.98 (t,J=4.8 Hz, 1H), 6.48 (d, J=6.8 Hz, 1H), 4.26 (t, J=5.5 Hz, 1H), 4.09 (qd,J=7.1, 3.1 Hz, 2H), 3.79 (t, J=5.6 Hz, 1H), 3.73 (br. s., 1H), 3.25 (s,3H), 3.05-3.22 (m, 1H), 1.89-2.14 (m, 4H), 1.21-1.37 (m, 4H), 1.18 (t,J=7.1 Hz, 3H) ppm. LC/MS: Calculated: 388.1; Found ES+ 389.1 (M+1) 391.1(M+1+2).

A 1 M solution of KOt-Bu in THF (0.20 eq) was added to a stirred mixtureof (13) (1.0 eq) in THF (8.0 vol) over 4 min at ambient temperature. Themixture was stirred for 2 h and quenched into a 9% aqueous KH₂PO₄solution (4.0 vol). IPAc (5 vol) was added, and the layers wereseparated. The organic layer was washed with 5% aqueous NaCl (4 vol) andconcentrated under reduced pressure with azeotropic removal of THF withIPAc. The solid was dissolved in IPAc (10 vol), passed through silicagel, eluted with IPAc, and concentrated under reduced pressure. Thesolids were dried at 20-25° C. under vacuum to afford (14) as a solid(70% yield). ¹H NMR (DMSO-d₆, 300 MHz): δ=7.70 (s, 1H), 7.57 (d, J=7.6Hz, 1H), 4.55-4.77 (m, 1H), 4.22-4.36 (m, 1H), 3.76-3.86 (m, 1H), 3.25(s, 3H), 3.04-3.19 (m, 1H), 2.33-2.47 (m, 2H), 1.98-2.20 (m, 2H), 1.61(d, J=11.1 Hz, 2H), 1.07-1.33 (m, 3H). LC/MS: Calculated: 342.1; found:ES+ (M+1) 343.0; (M+2+1) 345.1.

A mixture of 5-bromo-2-iodopyridine (1.0 eq) in DCM (12 vol) was cooledto −78° C. and treated with n-BuLi (2.5 M solution in hexanes, 1.0 eq).The mixture was treated with acetone-¹³C₃ (10 eq) while maintaining thetemperature below −55° C., cooled to −78° C., and held for 30 min. Thereaction mixture was warmed to −40° C. over 1 h, warmed to −15° C.,quenched with water (10 vol), warmed to 10° C. over 10 minutes, and thelayers were separated. The aqueous phase was extracted with DCM, and theorganic layers were washed with water, saturated aqueous NaCl, driedover Na₂SO₄, and filtered. The cake was washed with DCM, and thefiltrate was concentrated under reduced pressure to obtain an oil. Theoil was dissolved in DCM (12.0 vol), and DMAP (0.05 eq) and TEA (3.0 eq)were added. The solution was cooled to 0-5° C. and treated with TMSCl(2.5 eq) over 15 minutes keeping the temperature below 5° C. The mixturewas stirred for 1.5 h, quenched with 5% aqueous NaHCO₃ (6.5 vol)maintaining the temperature at 10-15° C. The layers were separated, andthe organic layer was washed with water and saturated aqueous NaCl. Theorganic layer was dried over Na₂SO₄, filtered, and concentrated underreduced pressure. Hexanes (2×9 vol) were charged and the mixture wasconcentrated under reduced pressure to afford an oil. The oil waspurified by column chromatography on silica gel (5% EtOAc in hexanes) toafford (15) (63% yield). ¹H NMR (MeOD, 300 MHz): δ=8.38 (d, J=2.1 Hz,1H), 7.78 (dd, J=8.6, 2.4 Hz, 1H), 7.48 (d, J=8.5 Hz, 1H), 1.61-1.70 (m,3H), 1.18-1.27 (m, 3H), 0.00 (s, 9H).

Compound (15) (1.0 eq), bis(pinacolato)diboron (1.0 eq) and KOAc (3.0eq) were stirred in 1,4-dioxane (8 vol) and treated with PdCl₂(dppf).DCMcomplex (0.015 eq). The mixture was heated to 90-95° C. and stirred for4.5 h. The reaction mixture was cooled to 20-25° C. over 1 h, dilutedwith MTBE (5 vol), filtered on a celite plug, and the cake was washedwith MTBE. The filtrate was washed with water, and the aqueous layer wasextracted with MTBE. The organic layers were washed with saturatedaqueous NaCl, dried over Na₂SO₄, and filtered. The filtrate wasconcentrated under reduced pressure to an oil, treated with MTBE andconcentrated to an oil three times. The oil was dried under high vacuumat 20-25° C. to afford a solid. This solid was dissolved in THF (7.5vol), treated with SiliaBond® Thiol (1× wt), stirred for 20 min,filtered, and the cake washed with THF. The filtrate was concentratedunder reduced pressure to afford a solid, which was dried under highvacuum. The solid was dissolved in MTBE, treated with silica gel (1×wt), and concentrated under reduced pressure. The silica gel containingthe crude product was purified by column chromatography on silica gel(eluent: MTBE) and concentrated under reduced pressure to obtain theproduct (16) as a solid (72% yield). ¹H NMR (CDCl₃, 300 MHz): δ=8.71 (s,1H), 7.90 (d, J=7.6 Hz, 1H), 7.45-7.55 (m, 1H), 1.64-1.72 (m, 3H), 1.25(d, J=4.0 Hz, 3H), 1.20 (s, 12H), 1.13 (s, 1H), 1.10 (s, 1H), 0.00 (s,9H) ppm. MS Calculated: 410.2, found ES+ 257 (as boric acid).

A slurry of compound (14) (1.0 eq) and compound (16) (1.20 eq) in IPA(10 vol) was treated with 2 M aqueous Na₂CO₃ (2.5 eq) and PdCl₂Amphos₂(0.0135 eq). The reaction mixture was heated to 70° C., stirred for 2 h,cooled to ambient temperature, and treated with EtOAc (38 vol) and water(13 vol). The organic layer was washed with 2% aqueous NaCl to reach pH6 and concentrated under reduced pressure. EtOAc (13 vol) was added tothe concentrate, the aqueous layer was extracted with EtOAc, and thecombined organic phases were concentrated under reduced pressure. Theresidue was dissolved in EtOAc and purified by column chromatography onsilica gel (EtOAc/hexanes), concentrated under reduced pressure andcooled to 0° C. The solids were dissolved in IPA, concentrated underreduced pressure, and dried under high vacuum to provide (17) as a solid(73% yield). ¹H NMR (DMSO-d₆, 300 MHz): δ=9.03 (d, J=1.9 Hz, 1H), 8.28(s, 1H), 8.25 (dd, J=8.4, 2.2 Hz, 1H), 7.68 (d, J=8.3 Hz, 1H), 7.61 (d,J=7.7 Hz, 1H), 4.81-4.99 (m, J=11.8, 7.9, 3.9, 3.9 Hz, 1H), 4.35 (d,J=6.2 Hz, 1H), 3.88 (d, J=6.4 Hz, 1H), 3.25-3.31 (m, 3H), 3.13-3.24 (m,1H), 2.52-2.67 (m, 2H), 2.13 (d, J=10.4 Hz, 2H), 1.79 (d, J=3.8 Hz, 3H),1.67 (d, J=10.6 Hz, 2H), 1.36 (d, J=4.0 Hz, 3H), 1.18-1.33 (m, 2H),0.06-0.18 (m, 9H). Calculated 402.2 (−TMS+1); ES+(M+2-TMS) 403.2.

A slurry of (17) (1.0 eq), ACN (10.0 vol) and water (2.5 vol) wastreated with 1 M HCl (0.185 eq) for 20 h and neutralized to pH 4-6 with1 M NaOH. The mixture was treated with water (50 vol) and EtOAc (75 vol)and the layers were separated. The aqueous layer was extracted withEtOAc and the combined organic layers were concentrated under reducedpressure. The residue was again treated with water (50 vol) and EtOAc(75 vol) and the layers were separated and the aqueous layer extractedwith additional EtOAc. The organic fractions were concentrated underreduced pressure with replacement of EtOAc by ACN addition. The residuewas dissolved in ACN (2.5 vol), and a small amount (0.02 eq) of thetarget product was added followed by additional ACN (0.8 vol). Thesolids were filtered, washed with ACN, and dried under a N₂ stream. Thesolid was dissolved in EtOAc and silica gel (1.9 wt) was added and themixture concentrated under reduced pressure. The silica gel containingthe crude product was purified by column chromatography on silica gel(eluent: EtOAc) and concentrated under reduced pressure with replacementof EtOAc by ACN addition. The material was dried under high vacuum,slurried in ACN (2.5 vol) for 20 h, and filtered to obtain (18) as asolid (34% yield). ¹H NMR (DMSO-d₆, 300 MHz): δ=9.02 (d, J=1.9 Hz, 1H),8.28 (s, 1H), 8.23 (dd, J=8.3, 2.1 Hz, 1H), 7.73 (d, J=8.5 Hz, 1H), 7.59(d, J=7.7 Hz, 1H), 5.24 (d, J=2.3 Hz, 1H), 4.80-5.00 (m, J=11.9, 8.0,3.9, 3.9 Hz, 1H), 4.36 (d, J=6.2 Hz, 1H), 3.88 (d, J=6.2 Hz, 1H),3.25-3.31 (m, 3H), 3.14-3.25 (m, 1H), 2.53-2.67 (m, 2H), 2.14 (d, J=10.4Hz, 2H), 1.68 (d, J=4.0 Hz, 5H), 1.18-1.35 (m, 5H). ¹³C NMR (DMSO-d₆, 75MHz): δ=168.7, 168.0, 167.0, 166.6, 166.3, 165.7, 164.9, 162.9, 162.1,157.4, 156.9, 156.4, 155.9, 154.9, 145.7, 145.7, 145.0, 137.0, 135.6,133.6, 133.3, 131.3, 119.9, 119.8, 86.4, 85.6, 79.2, 76.5, 75.7, 74.3,74.0, 73.8, 73.5, 73.2, 73.0, 56.3, 53.3, 47.6, 47.3, 47.0, 41.6, 41.3,41.0, 40.7, 40.5, 40.2, 39.9, 32.5, 32.1, 31.8, 31.6, 31.0, 27.1.Calculated 402.2, found ES+(M+1) 403.2.

6.3.3 Synthesis of ¹³C Enriched Metabolite of Compound A

¹³C₅-labeled metabolite of Compound A was prepared as follows.

A slurry of 3,5-dibromopyrazin-2-amine (1 eq) in acetone (10 vol) wastreated with K₂CO₃ (0.8× wt) and ethyl bromoacetate-¹³C₂ (0.87× wt) wereadded, and the mixture was heated to 30° C. Bu₄NHSO₄ (0.1× wt) wasadded, and the mixture was stirred for 46 h at reflux. Additional ethylbromoacetate-¹³C₂ was added in portions and the mixture was held atreflux until completion was achieved (˜24 h). The reaction mixture wascooled to 20-25° C., filtered, and the filter cake was washed twice withacetone. The filtrate was concentrated under reduced pressure, dissolvedin EtOAc, washed twice with water and then with a 5% aqueous NaCl. Thecombined aqueous washes were extracted with EtOAc, and the combinedorganic fractions were treated with MgSO₄ (0.3× wt) and Ecosorb C-906(0.1× wt) for 13 h at 30° C. The mixture was cooled to 20° C. andfiltered. The collected solids were washed twice with EtOAc, and thefiltrate was concentrated to a solid which was dissolved in EtOAc (0.9vol) and treated with heptane (5.7 volumes) over 40 min at 20-25° C. Thesuspension was stirred for 4 h and filtered. The isolated solids werewashed with heptane and dried under reduced pressure at 35-40° C. toprovide 11.8 g of (12) as a solid (46% yield). LC/MS: Calculated [M+1]342.3; Observed 342, 344.

A slurry of compound 12 (1 eq) and trans-4-aminocyclohexanolhydrochloride (1.5 eq) in NMP (5 vol) at ambient temperature was treatedwith DIPEA (3.5 eq). The mixture was heated to 125-130° C. and held for18 h. The solution was cooled to 20-25° C., treated with EtOAc (10 vol),and washed three times with 5% aqueous NaCl and once with water. Thesolution was concentrated under reduced pressure to 2 vol and the slurrywas stirred for 18 h at ambient temperature. The solids were collectedby filtration and dried to obtain compound (19) (24% yield). Thefiltrate was concentrated under reduced pressure, stirred for 18 h atambient temperature, treated with EtOAc (1-2 vol) and filtered. Thesolids were dried under reduced pressure to obtain compound (19) (14%yield). LC/MS: Calculated [M+1] 375, 377; Observed 375, 377.

Compound (19) (1× wt) and a 21% H₃PO₄ solution (10 vol) were combined atambient temperature and heated to 75-80° C. and stirred for 16 h. Thebatch was cooled to 20-25° C. and then filtered, and the filter cake waswashed with water. The solid was suspended in water (10 vol) and stirredfor 2 h at 20-25° C. The product was filtered, washed twice with water,and dried under reduced pressure at 45-50° C. (20) as a solid (65%yield). LC/MS: Calculated [M+1] 329, 331; Observed 329, 331.

5-bromo-2-iodopyridine (1.0 eq) in DCM (12 vol) was cooled to −78° C.and treated with n-BuLi (1.4 vol of 2.5 M in hexanes) over 45 min. After40 min, ¹³C₃-acetone (2.0 eq) was added over 50 minutes keeping thereaction mixture below −70° C. The mixture was stirred for 2 h below−70° C., warmed to −14° C. over 2 h, quenched with water (10 vol)between −15° and 10° C., and warmed to 10° C. The aqueous layer wasextracted with DCM, and the combined organic layers were washed withwater and saturated aqueous NaCl, dried over MgSO₄, filtered, and washedwith DCM. The filtrate was concentrated under reduced pressure to obtain(21) as a liquid (62% yield). LC/MS: Calculated [M+1] 219, 221; Observed219, 221.

A solution of compound (21) (1 eq) in DCM (395 mL) was treated with DMAP(0.01 eq) and the solution was cooled to 0° C. TEA (1 eq) and TMSCl (1.5eq) were added and the reaction mixture was stirred at 0-5° C. for 2 h,quenched by addition of saturated aqueous NaHCO3 (2.3 vol) and water(2.3 vol). DCM was added and the layers were separated. The organiclayer was washed with water and saturated aqueous NaCl, dried overMgSO4, and filtered. The cake was rinsed with DCM and the filtrate wasconcentrated under reduced pressure, treated with hexanes, andconcentrated under reduce pressure to obtain crude (15). The crudeproduct was purified by column chromatography on silica gel (eluent: 5%EtOAc in hexanes) and concentrated to a residue. The residue was treatedwith hexanes and concentrated to an oil to provide compound (15) as anoil (61% yield). LC/MS: Calculated [M+1] 291, 293; Observed 291, 293. ¹HNMR (CDCl₃, 300 MHz): δ=8.39 (d, J=2.1 Hz, 1H), 7.61 (dd, J=2.3, 8.5 Hz,1H), 7.41 (d, J=8.5 Hz, 1H), 1.57-1.73 and 1.17-1.28 (2 m, 6H, 13CH3),0.00 (s, 9H). ¹³C NMR (CDCl₃, 75 MHz) δ=164.63 (d, JC-C=6 Hz), 146.57(d, JC-C=6 Hz), 136.44 (d, JC-C=2 Hz), 118.48 (d, JC-C=4 Hz), 115.94,74.59 (t, JC-C=39 Hz), 28.80 (d, JC-C=39 Hz), 0.50.

A solution of compound (15) (1 eq) in 1,4-dioxane (8 vol) was treatedwith KOAc (2.2 eq), bis(pinacolato)diboron (1 eq), and PdCl₂(dppf).DCMcomplex (0.02 eq). The contents were heated to reflux, held for 4 h,cooled to ambient temperature and treated with MTBE (10 vol). The slurrywas filtered and the filter cake was washed with MTBE. The filtrate waspassed through a 0.45 mm filter, transferred to a separatory funnel, andwashed with water. The aqueous phase was extracted with MTBE and treatedwith aqueous NaCl. The combined organic extracts were washed withsaturated aqueous NaCl, dried over MgSO₄, and filtered. The filtrate wasconcentrated under reduced pressure. The residue was dissolved in ACN(1.1 vol) at 45° C., and transferred with ACN (3.9 vol) to a flask. Thecrude product was heated to 40-50° C., cooled to ambient temperature,agitated for 14.5 h, cooled to 0-5° C., and stirred for 2 h. The productwas filtered, washed with cold ACN, and dried under vacuum at 40-55° C.to provide (16) as a solid (65% yield). ¹H NMR (DMSO-d₆, 300 MHz):δ=9.03 (d, J=1.9 Hz, 1H), 8.28 (s, 1H), 8.25 (dd, J=8.4, 2.2 Hz, 1H),7.68 (d, J=8.3 Hz, 1H), 7.61 (d, J=7.7 Hz, 1H), 4.81-4.99 (m, J=11.8,7.9, 3.9, 3.9 Hz, 1H), 4.35 (d, J=6.2 Hz, 1H), 3.88 (d, J=6.4 Hz, 1H),3.25-3.31 (m, 3H), 3.13-3.24 (m, 1H), 2.52-2.67 (m, 2H), 2.13 (d, J=10.4Hz, 2H), 1.79 (d, J=3.8 Hz, 3H), 1.67 (d, J=10.6 Hz, 2H), 1.36 (d, J=4.0Hz, 3H), 1.18-1.33 (m, 2H), 0.06-0.18 (m, 9H). ¹H NMR (CDCl₃, 300 MHz):δ=8.71 (s, 1H), 7.89 (dd, J=0.8, 7.9 Hz, 1H), 7.49 (d, J=7.7 Hz, 1H),1.61-1.75 and 1.23-1.32 (2 m, 6H, 13CH3), 1.21 (s, 12H), 0.00 (s, 9H).¹³C NMR (CDCl₃, 75 MHz) δ=168.76, 151.71 (d, JC-C=6 Hz), 140.21, 115.97(d, JC-C=4 Hz), 81.55, 74.69 (t, JC-C=39 Hz), 28.60 (d, JC-C=39 Hz),22.41, 0.087. LC/MS: LC/MS: Calculated [M+1] 339.2; Observed 257.2 (asboric acid).

A solution of (16) (1 eq) in 1,4-dioxane (4 vol) was cooled to 15-20° C.and treated with 4 M HCl in 1,4-dioxane (2.1 eq). The slurry was treatedwith heptane (3.75 vol), cooled to 0-5° C., stirred for 1-2 h, andfiltered. The product was washed with heptane and dried under vacuum at50-60° C. to obtain (22) as a solid (94% yield). ¹H NMR (CDCl₃, 300MHz): δ=16.56 (br. s., 1H), 9.05 (s, 1H), 8.54 (d, J=7.9 Hz, 1H), 7.78(dd, J=1.4, 8.0 Hz, 1H), 4.1-6.3 (br. s., 1H), 1.95-1.98 and 1.52-1.56(2 m, 6H, 13CH3), 1.30 (s, 12H). ¹³C NMR (CDCl₃, 75 MHz): δ=164.79 (d,JC-C=47 Hz), 150.91 (d, JC-C=2.4 Hz), 146.50, 122.26 (d, JC-C=2.8 Hz),85.66, 71.92 (t, JC-C=38 Hz), 29.89 (d, JC-C=38 Hz), 24.83. LC/MS:Calculated [M+1] 303; Observed 185 (as boric acid).

Compound (20) (1 eq), Compound (22) (1.1 eq), PdCl₂Amphos₂ (0.009 eq),and THF (5 vol) were combined and treated with a solution of K₂CO₃ (2.1eq) in water (3.75 vol). The mixture was heated to reflux, held for 6 h,cooled to ambient temperature, stirred for 11 h, and filtered. Thefilter cake was washed twice with 1 vol of THF/water (5:8) and thefiltrate was diluted with THF (6.75 vol). The filtrate was heated to40-45° C. and treated with toluene (6.75 vol). The organic layer waswashed with a solution of KH₂PO₄ in water (0.04 w/w) and the layers wereseparated. The organic layer was heated to 40-45° C. and treated withSiliaBond® Thiol for 2 h. The slurry was cooled to ambient temperature,filtered, and the filter cake was washed with THF. The filtrate wastreated with activated carbon (decolorizing) for 4 h at ambienttemperature, filtered, and the filter cake was washed with THF. Thefiltrate was concentrated under reduced pressure, dissolved in DCM, andconcentrated under reduce pressure. The residue was dried under vacuum,treated with THF, heated to 40-45° C., and treated with silica gel. Theslurry was concentrated under reduced pressure and the silica gelcontaining the crude product was purified by column chromatography onsilica gel (eluent 0-41% THF in DCM), concentrated under reducedpressure, and dried under vacuum at 30-40° C. to obtain crude (23). Thecrude (23) and BHT (0.0005× wt) were treated with IPA/water (1:1.65),heated to 60° C., held for 1 h, cooled to ambient temperature, and heldfor 16 h. The slurry was heated to 50-60° C., treated with IPA (0.8 vol)and water (23 vol). The slurry was cooled to ambient temperature andfiltered. The product was washed with IPA/water (10:90) and dried undervacuum at 50-60° C. to afford (23) as a solid (85% yield). ¹H NMR(CDCl₃, 300 MHz): δ=9.03 (d, J=1.9 Hz, 1H), 8.27 (s, 1H), 8.23 (dd,J=2.1, 8.3 Hz, 1H), 7.72 (d, J=8.3 Hz, 1H), 7.59 (d, J=7.6 Hz, 1H), 5.23(m, 1H), 4.81-4.92 (m, 1H), 4.65 (d, J=4.3 Hz, 1H), 4.36 (d, J=6.4 Hz,1H), 3.88 (d, J=6.2 Hz, 1H), 3.41-3.57 (m, 1H), 2.53-2.71 (m, 2H), 1.95(d, J=10.4 Hz, 2H), 1.66-1.69 and 1.24-1.27 (2 m, 6H, 13CH3), 1.29-1.37(m, 2H). ¹³C NMR (CDCl₃, 75 MHz): δ=165.34 (d, JC-C=52 Hz), 144.46 (d,JC-C=5.6 Hz), 143.74 (d, JC-C=2 Hz), 135.78, 134.28, 132.28, 132.01,130.02, 118.54 (d, JC-C=42 Hz), 72.18 (d, JC-C=38 Hz), 68.54, 52.03,45.85 (d, JC-C=52 Hz), 35.09, 30.53, (d, JC-C=39 Hz), 26.11. LC/MS:Calculated [M+1] 388; Observed 389.

6.3.4 Synthesis of ²H Enriched Compound A

Deuterium-enriched Compound A can be prepared as follows.

Compound 24 can be made using the route above wherein all of theexchangeable protons are replaced with deuterium. Starting with CompoundA, the acidic protons can be exchanged in the presence of base (such assodium tert-butoxide, potassium carbonate and1,8-diazabicyclo[5,4,0]undec-7-ene) and a deuterium source (such astert-BuOD, MeOD, EtOD, iPrOD, AcOD, D₂O) to give Compound 24. A solvent(such as tetrahydrofuran, dimethylformamide, or dimethylsulfoxide) canbe used to facilitate the reaction. The hydrogen isotopes on the alcoholand the secondary amine could be either hydrogen or deuterium dependingon the workup. A workup solvent with an exchangeable proton (such asH₂O, MeOH or EtOH) will provide 25, while a workup solvent with anexchangeable deuterium (e.g., D₂O, MeOD, EtOD) will afford 24.

For example, Compound A (10 g, 25.2 mmol) was treated with K₂CO₃ (3.48g, 25.2 mmol) in 20% THF/D20 at 50-60° C. for 15 h. After cooling toroom temperature, the mixture was extracted with 2-Me-THF, and theorganic layer was washed 3 times with water to allow proton exchange ofthe alcohol and the pyrazine groups. The organic layer was concentratedto a crude oil and crystallization from IPA/water to afford Compound 25(7.6 g, 76%) as an off-white solid; ¹H NMR (300 MHz, CDCl₃) δ 9.02 (d,J=1.5 Hz, 1H), 8.27-8.05 (m, 2H), 7.49 (d, J=8.3 Hz, 1H), 5.51 (s, 1H),5.15-4.97 (m, 1H), 4.93 (s, 1H), 3.40 (s, 3H), 3.37-3.23 (m, 1H),2.79-2.53 (m, 2H), 2.43-2.11 (m, 2H), 1.92-1.70 (m, 2H), 1.60 (s, 6H),1.52-1.29 (m, 2H); ¹³C NMR (300 MHz, CDCl₃) δ 165.6, 164.8, 144.6,143.1, 136.7, 136.5, 133.6, 132.0, 130.8, 118.7, 78.5, 71.9, 55.9, 53.2,46.4, 31.6, 30.6, 26.4; LCMS (EI) m/z calcd. for C₂₁H₂₅D₂N₅O₃ [M+H]⁺,400.2; found 400.2

6.3.5 Synthesis of ²H Enriched Metabolite of Compound A

A deuterium-enriched metabolite of Compound A can be prepared asfollows.

Compound 26 can be made using the route above wherein all of theexchangeable protons are replaced with deuterium. Starting with Compound6, the acidic protons can be exchanged in the presence of base (such assodium tert-butoxide, potassium carbonate and1,8-diazabicyclo[5,4,0]undec-7-ene) and a deuterium source (such astert-BuOD, MeOD, EtOD, iPrOD, AcOD, D20) to give Compound 26. A solvent(such as tetrahydrofuran, dimethylformamide, or dimethylsulfoxide) canbe used to facilitate the reaction. The hydrogen isotopes on the twoalcohols and the secondary amine could be either hydrogen or deuteriumdepending on the workup. A workup solvent with an exchangeable proton(such as H₂O, MeOH or EtOH) will provide 27, while a workup solvent withan exchangeable deuterium (e.g., D₂O, MeOD, EtOD) will afford 26.

6.4 Pharmaceutical Compositions

6.4.1 Tablets

Compound A was formulated as tablets containing about 5 mg, 20 mg, and50 mg of Compound A as an active pharmaceutical ingredient. Theexcipients and carriers that were used in the tablet formulations aresummarized in Table 6, along with their intended functions.

TABLE 6 Pharmaceutical Acceptable Excipients and Carriers IngredientsFunction Lactose monohydrate, NF (Fast Flo 316) Diluent Microcrystallinecellulose, NF (Avicel pH 101) Diluent/binder Microcrystalline cellulose,NF (Avicel pH 102) Diluent/binder Corn starch, NF Disintegrant/lubricantPregelatinized starch, NF (Starch 1500) Binder/Disintegrant Lactoseanhydrous, NF Diluent Croscarmellose sodium, NF (Ac-Di-Sol) DisintegrantStearic acid, NF Lubricant Magnesium Stearate, NF Lubricant

General Method for Tablet Preparation.

Tablets were produced at batch size ranging from 0.5 to 2.2 kg. Form Aof compound A was first mixed/blended with binders, diluent(s), and/ordisintegrant (e.g., lactose monohydrate (NF), croscarmellose sodium(NF), and/or microcrystalline cellulose (NF)) using a Globepharma 4-8quart Bin Blender. The mixture was then sieved via 18 mesh screen. Thesieved mixture was further mixed/blended with a Globepharma 4-8 quartBin Blender. After lubricant(s) (e.g., stearic acid (NF) and/ormagnesium stearate (NF)) were sieved via 30 mesh screen, thelubricant(s) were then added to the mixture. The resulting mixture wasthen mixed/blended with a Globepharma 4-8 quart Bin Blender. The mixturewas then compressed into tablets with a rotary table press, and thencoated in an Ohara 8″ pan. The tablets thus produced were evaluated fortheir powder characteristics, tablet characteristics, drug productphotostability/short term stability, and manufacturing process.

Tablet formulations I to VIII of Compound A are summarized in Table 7 toTable 14. The process parameters for tablet preparation(blending/compression) are summarized in Table 15 and Table 16. It wasobserved that the tablets of Formulations I to VIII showeddiscoloration. Picking was observed when compressing Formulations I toIV. The addition of stearic acid in Formulations V to VIII improvedlubrication without impacting disintegration and compressibility.Compressibility of Formulation II was not acceptable when replacinglactose by pregelatinized starch and tablet hardness could not exceed4.1 kp (average). Lactose monohydrate, NF (Fast Flo 316) was used as analternate diluent and was preferred over lactose anhydrous (FormulationIII) for its flowability properties. Both Avicel PH 101 and PH 102 weretested for binding properties (Formulations III and IV). Avicel PH 102'slarger particle size, and more spherical particle shape provided betterflow than Avicel PH 101.

TABLE 7 Tablet Formulation I Amounts Ingredients mg % Compound A 50.016.7 Lactose monohydrate, NF (Fast Flo 316) 145.1 48.3 Microcrystallinecellulose, NF (Avicel pH 101) 93.1 31.0 Croscarmellose sodium, NF(Ac-Di-Sol) 9.0 3.0 Magnesium Stearate, NF 3.0 1.0 Total 300.0 100

TABLE 8 Tablet Formulation II Amounts Ingredients mg % Compound A 50.016.7 Lactose monohydrate, NF (Fast Flo 316) 168.0 56.0 Pregelatinizedstarch, NF (Starch 1500) 70.1 23.3 Croscarmellose sodium, NF (Ac-Di-Sol)9.0 3.0 Magnesium Stearate, NF 3.0 1.0 Total 300.0 100

TABLE 9 Tablet Formulation III Amounts Ingredients mg % Compound A 50.016.7 Lactose anhydrous, NF 145.1 48.3 Microcrystalline cellulose, NF(Avicel pH 101) 93.1 31.0 Croscarmellose sodium, NF (Ac-Di-Sol) 9.0 3.0Magnesium Stearate, NF 3.0 1.0 Total 300.0 100

TABLE 10 Tablet Formulation IV Amounts Ingredients mg % Compound A 50.016.7 Lactose monohydrate, NF (Fast Flo 316) 145.0 48.3 Microcrystallinecellulose, NF (Avicel pH 102) 93.0 31.0 Croscarmellose sodium, NF(Ac-Di-Sol) 9.0 3.0 Magnesium Stearate, NF 3.0 1.0 Total 300.0 100

TABLE 11 Tablet Formulation V Amounts Ingredients mg % Compound A 50.011.9 Lactose monohydrate, NF (Fast Flo 316) 220.48 52.5 Microcrystallinecellulose, NF (Avicel pH 102) 130.20 31.0 Croscarmellose sodium, NF(Ac-Di-Sol) 12.6 3.0 Stearic acid, NF 2.52 0.6 Magnesium Stearate, NF4.20 1.0 Total 420.0 100

TABLE 12 Tablet Formulation VI Amounts Ingredients mg % Compound A 50.011.9 Lactose monohydrate, NF (Fast Flo 316) 182.20 63.1 Microcrystallinecellulose, NF (Avicel pH 102) 54.0 18.0 Croscarmellose sodium, NF(Ac-Di-Sol) 9.0 3.0 Stearic acid, NF 1.80 3.0 Magnesium Stearate, NF 3.01.0 Total 300.0 100

TABLE 13 Tablet Formulation VII Amounts Ingredients Mg % Compound A 50.016.7 Lactose monohydrate, NF (Fast Flo 316) 265.0 88.3 Microcrystallinecellulose, NF (Avicel pH 102) 75.60 25.2 Corn starch, NF 12.6 4.2Croscarmellose sodium, NF (Ac-Di-Sol) 12.6 4.2 Magnesium Stearate, NF4.20 1.4 Total 420.0 100

TABLE 14 Tablet Formulation VIII Amounts Ingredients Mg % Compound A50.0 16.7 Lactose monohydrate, NF (Fast Flo 316) 136.0 45.3Microcrystalline cellulose, NF (Avicel pH 102) 93.0 31.0 Corn starch, NF9.0 3.0 Croscarmellose sodium, NF (Ac-Di-Sol) 9.0 3.0 MagnesiumStearate, NF 3.0 1.0 Total 300.0 100

TABLE 15 Tablet Process Parameters Equipment/Process Parameters I II IIIIV Batch size (kg) 0.5 0.5 0.5 0.5 Bin blender (quart) 4 4 4 4Pre-blending time (min) 20/10 20/10 20/10 20/10 Lubrication time (min) 33 3 3 Actual weight (mg) 299 301 307 297 291-309 295-310 301-311 290-300Bulk density (g/cc) 0.4 0.53 0.37 0.42 Tooling (round, SC) 12/32 12/3212/32 12/32 Hardness (average in Kp) 7.9 4.1 7.9 7.4 Thickness (averagein mm) 3.95 3.86 3.98 3.86 Friability (4 min) (%) 0 0.1 0 0.1Disintegration time (max) (sec) 18 75 55 21 Observation Picking PickingPicking Picking

TABLE 16 Tablet Process Parameters Equipment/Process Parameters V VI VIIVIII Batch size (kg) 0.5 0.5 0.5 0.5 Bin blender used (quart) 4 4 4 4Pre-blending time (min) 20/10 20/10 20/10 20/10 Lubrication time (min) 33 3 3 Actual weight (mg) 418 299 419 301 413-421 293-307 413-426 296-305Bulk density (g/cc) 0.45 0.43 0.48 0.43 Tooling (round, SC) 12/32 12/3212/32 12/32 Hardness (average in Kp) 9.1 8.5 9.0 8.4 Thickness (averagein mm) 5.20 3.8 4.12 3.86 Friability (4 min) (%) 0.3 0.2 0.2 0.1Disintegration time (max) (sec) 31 30 29 20 Observation None None NoneNone

Tablet formulations IX to XI of Compound A are summarized in Table 17 toTable 19. The process parameters for their preparation are summarized inTable 20 and Table 21.

TABLE 17 Tablet Formulation IX Amounts Ingredients mg % Compound A 50.015.4 Lactose monohydrate, NF (Fast Flo 316) 151.5 46.6 Microcrystallinecellulose, NF (Avicel pH 102) 100.75 31.0 Corn starch, NF 9.75 3.0Croscarmellose sodium, NF (Ac-Di-Sol) 9.75 3.0 Magnesium Stearate, NF3.25 1.0 Total 325.0 100 Opadry pink 03K140004 4% weight gain

TABLE 18 Tablet Formulation X Amounts Ingredients mg % Compound A 50.015.4 Lactose monohydrate, NF (Fast Flo 316) 149.55 46.0 Microcrystallinecellulose, NF (Avicel pH 102) 100.75 31.0 Corn starch, NF 9.75 3.0Croscarmellose sodium, NF (Ac-Di-Sol) 9.75 3.0 Stearic acid, NF 1.95 0.6Magnesium Stearate, NF 3.25 1.0 Total 325.0 100 Opadry pink 03K140004 4%weight gain

TABLE 19 Tablet Formulation XI Amounts Ingredients mg % Compound A 5.03.85 Lactose monohydrate, NF (Fast Flo 316) 74.82 57.55 Microcrystallinecellulose, NF (Avicel pH 102) 40.30 31.00 Corn starch, NF 3.90 3.00Croscarmellose sodium, NF (Ac-Di-Sol) 3.90 3.00 Stearic acid, NF 0.780.60 Magnesium Stearate, NF 1.30 1.00 Total 130.0 100 Opadry beige03K170001 4% weight gain

TABLE 20 Tablet Process Parameters Equipment/Process ParametersBlending/Compression IX X XI Batch size (kg) 0.65 0.65 0.52 Bin blenderused (quart) 4 4 4 Pre-blending time (min) 20/10 20/10 20/10 Lubricationtime (min) 3 3 3 Actual weight (mg) 323 326 131 318-328 316-333 130-134Bulk density (g/cc) 0.40 0.42 0.48 Tooling (round, SC) 12/32 12/32 ¼Hardness (average in Kp) 9.3 9.1 5.9 Thickness (average in mm) 4.09 4.123.72 Friability (4 min) (%) 0.1 0.1 0.1 Disintegration time (max) (sec)39 27 24 Observations Picking None None

TABLE 21 Tablet Process Parameters Equipment/Process Parameters CoatingIX X XI Batch size (kg) 0.27 0.27 0.30 Weight gain (%) 4 4 4 Solid insuspension (%) 12 12 12 Pan (inch) 8 8 8 Nozzle size (mm) 0.8 0.8 0.8Atomizing air pressure (PSI)  9-10 10-12  9-10 Pattern (PSI) 12-13 12-1311-12 Distance gun-ben (inch) 3 3 3 Airflow (CFM) 75 75 75 Pan speed(RPM) 16-18 14-17 14-17 Inlet temperature (° C.) 75 75 72-73 Exhausttemperature (° C.) 51-53 51-53 49-50 Spray rate 5-7 4-6 4-6 ObservationAcceptable appearance

The 5 mg and 50 mg tablets (core and coated) were subjected to shortterm stability and photo-stability evaluations. The short term stabilityof the 50 mg tablets was tested by storing for 2 weeks at 40° C./75% RHin an open bottle. The results are summarized in Table 22.

TABLE 22 Tablet Formulation X (50 mg) Tablet Short Term StabilityCompound A (%) Total Impurities (%) After 2 wks at After 2 wks at TabletInitial 40° C./75% RH Initial 40° C./75% RH Core 99.5 98.7 0.29 0.54Coated 100.1 99.9 0.25 0.29

The photo-stability of the 50 mg tablets was also tested and the resultsare summarized in Table 23.

TABLE 23 Tablet Formulation X (50 mg) Tablet Photo-Stability Compound A(%) Total Impurities (%) Photo-stability Photo-stability Tablet ControlSample Control Sample Core 99.3 99.0 0.21 1.25 Coated 99.6 97.4 0.260.31

The short term stability of the 5 mg tablets was tested by storing themfor 2 weeks at 40° C./75% RH in an open bottle. The results aresummarized in Table 24. No major increase of impurity was observed forthe 50 mg coated tablets after two weeks at 40° C./75% RH and lightexposure. The coating appears to offer acceptable protection againstmoisture and light.

TABLE 24 Tablet Formulation X (5 mg) Tablet Short Term StabilityCompound A (%) Total Impurities (%) After 2 wks at After 2 wks at TabletInitial 40° C./75% RH Initial 40° C./75% RH Core 102.3 102.3 0.24 0.92Coated 101.1 100.7 0.21 1.11

The photo-stability of the 5 mg tablets was a so tested an t e resultsare summarized in Table 25.

TABLE 25 Tablet (5 mg) Tablet Photo-Stability Compound A (%) TotalImpurities (%) Photo-stability Photo-stability Tablet Control SampleControl Sample Core 99.5 97.9 0.27 2.85 Coated 99.0 101.0 0.23 0.84

Tablet formulations XII (50 mg), XIII (20 mg), and XIV (5 mg) aresummarized in Table 26, Table 27 and Table 28.

TABLE 26 Tablet Formulation XII (50 mg) Amounts Ingredients mg %Compound A 50.0 15.38 Lactose monohydrate, NF (Fast Flo 316) 159.9549.22 Microcrystalline cellulose, NF (Avicel pH 102) 100.75 31.00Croscarmellose sodium, NF (Ac-Di-Sol) 9.75 3.00 Stearic acid, NF 1.300.40 Magnesium Stearate, NF 3.25 1.00 Total 325.0 100 Opadry pink03K140004 13.0 4% weight gain

TABLE 27 Tablet Formulation XIII (20 mg) Amounts Ingredients mg %Compound A 20.0 15.38 Lactose monohydrate, NF (Fast Flo 316) 63.98 49.22Microcrystalline cellulose, NF (Avicel pH 102) 40.30 31.00Croscarmellose sodium, NF (Ac-Di-Sol) 3.90 3.00 Stearic acid, NF 0.520.40 Magnesium Stearate, NF 1.30 1.00 Total 130.0 100 Opadry yellow03K12429 5.2 4% weight gain

TABLE 28 Tablet Formulation XIV (5 mg) Amounts Ingredients mg % CompoundA 5.0 3.80 Lactose monohydrate, NF (Fast Flo 316) 78.98 60.70Microcrystalline cellulose, NF (Avicel pH 102) 40.30 31.00Croscarmellose sodium, NF (Ac-Di-Sol) 3.90 3.00 Stearic acid, NF 0.520.40 Magnesium Stearate, NF 1.30 1.00 Total 130.0 100 Opadry II pink85F94211 5.2 4% weight gain

No event was observed during the preparation of the tablets ofFormulations XII, XIII, or XIV. The 20 mg and 50 mg tablets werecompressed at various compression forces to assess compressibility anddefine a hardness range. The parameters for the preparation of thetablets to assess compressibility are summarized in Table 29(blending/compression) and Table 30 (coating). The 20 mg tablets werecoated with Opadry Yellow 03K12429, whereas the 50 mg tablets were notcoated. The core and coated tablets (20 mg) were tested for dissolution.It was found that there is no significant difference between thedissolution of the core and coated tablets (FIG. 5).

TABLE 29 Process Parameters for 50 mg and 20 mg Tablet Formulations(Blending/Compression) Equipment/Process Parameter 50 mg 20 mg BatchSize (kg) 2.21 (Common Blend) Bin Blende used (quart) 8 Pre-blendingtime (min) 20/10 Lubrication time (min) 3 3 Actual weight (mg) 327313-339 129 124-135 Bulk density (g/cc) 0.41 0.41 Tooling (round, SC)12/32 ¼ Hardness (average in Kp) Hig High-13.6 High-9.0 Low-5.9 Low-3.8Target-9.9 Target-6.1 Thickness (average in mm) 4.26 3.76 Friability (4min) (%) 0.09 0.04 Disinegration time (max) (sec) 39 22 ObservationsNone None

TABLE 30 Process Parameters for Formulation XIII (Coating)Equipment/Process Parameter 20 mg Batch size (kg) 0.27 Weight gain (%) 4Solid in suspension (%) 12 Pan (inch) 8 Nozzle size (mm) 0.8 Atomizingair pressure (PSI)  9-10 Pattern (PSI) 11-12 Distance gun-bed (inch) 3Airflow (CFM) 75 Pan speed (RPM) 14-16 Inlet temperature (° C.) 65Exhaust temperature (° C.) 45-47 Spray rate 4-5 Observation Acceptablecoating

Batch formulations of Compound A are summarized in Table 31.

TABLE 31 Batch Tablet Formulations 5 mg 20 mg Ingredients grams gramsCompound A 45.0 180.0 Lactose monohydrate 710.82 575.82 Microcrystallinecellulose 362.70 362.70 Croscarmellose sodium 35.10 35.10 Stearic acid4.68 4.68 Magnesium stearate 11.70 11.70 Total 1170.0 1170.0 Opadry ® IIPink 65.52 — Opadry ® Yellow — 65.52

Tablet formulation XV (45 mg) is summarized in Table 32. Tabletformulation XV can be prepared using methodology provided herein orother methods known to one skilled in the art.

TABLE 32 Tablet Formulation XV (45 mg) Amounts Ingredients mg % CompoundA 45.0 15.38 Lactose monohydrate, NF (Fast Flo 316) 143.955 49.22Microcrystalline cellulose, NF (Avicel pH 102) 90.675 31.00Croscarmellose sodium, NF (Ac-Di-Sol) 8.775 3.00 Stearic acid, NF 1.1700.40 Magnesium Stearate, NF 2.925 1.00 Total 292.50 100 Opadry pink03K140004 11.7 4.0% weight gain

The batch size of the current 45 mg strength tablet is approx. 10,000tablets or approximately 3.5 kg (approximately 20% overage is dispensedto allow for losses during manufacturing).

No picking or sticking was visually observed during the preparation ofthe tablets of Formulations XIII or XIV. As a result, stearic acid wasremoved from the tablet formulation.

Additionally, Compound A is susceptible to hydrolysis. Accordingly,without being limited by theory, a low-moisture grade microcrystallinecellulose (Avicel pH 112) was used in place of Avicel pH 102 to minimizeor prevent hydrolysis.

Tablet formulations XVI (15 mg) and XVII (30 mg) are summarized in Table33 and Table 34, below. Tablet formulations XVI and XVII can be preparedusing blending/sieving via a Comil process. After lubrication, themixture is then compressed into tablets and film-coated.

TABLE 33 Tablet Formulation XVI (15 mg) Amounts Ingredients mg %Compound A 15.0 15.38 Lactose monohydrate, NF (Fast Flo 316) 48.37 49.62Microcrystalline cellulose, NF (Avicel pH 112) 30.23 31.00Croscarmellose sodium, NF (Ac-Di-Sol) 2.925 3.00 Magnesium Stearate, NF0.975 1.00 Total 97.50 100 Opadry II pink 85F94211 3.9 4% weight gain

TABLE 34 Tablet Formulation XVII (30 mg) Amounts Ingredients mg %Compound A 30.0 15.38 Lactose monohydrate, NF (Fast Flo 316) 96.75 49.62Microcrystalline cellulose, NF (Avicel pH 112) 60.45 31.00Croscarmellose sodium, NF (Ac-Di-Sol) 5.85 3.00 Magnesium Stearate, NF1.95 1.00 Total 195.0 100 Opadry pink 03K140004 7.8 4% weight gain

6.4.2 Development of an Oral Dose Vehicle of ¹⁴C Enriched Compound A

A solution was prepared using appropriate amounts of 50:50 (v:v)EtOH:PEG 400, [¹⁴C]-Compound A, and Compound A to achieve a finalconcentration of 28.6 mg/mL. An aliquot of the solution was transferredto a white Size 00 Capsugel® V Caps Plus Hypromellose capsule for doseadministration. Preliminary stability data indicated that the in-processbulk solution is stable for at least 48 hours when stored atrefrigerated conditions and protected from light.

Compound A drug substance was dissolved in five different solventcombinations of EtOH and PEG 400. The solvent combinations selected were100% EtOH, 80:20 (v:v) EtOH:PEG 400, 50:50 (v:v) EtOH:PEG 400, 20:80(v:v) EtOH:PEG 400 and 100% PEG 400. Due to solubility and viscosityissues, the 100% EtOH and 100% PEG 400 formulations were not analyzed.

The 80:20 (v:v) EtOH:PEG 400, 50:50 (v:v) EtOH:PEG 400 and 20:80 (v:v)EtOH:PEG 400 solutions were prepared at a concentration of 28.6 mg/mLand diluted to 257 μg/mL for analysis. These samples were analyzed atT=0 and stored at RTmp/PFL and REF/PFL until analysis at T=72 hourspost-preparation.

Solution stability was performed on the final [¹⁴C]-Compound A dosingsolution to establish stability for at least 48 hours protected fromlight at refrigerated and room temperature conditions. Followinganalysis at T=0, T=24 hours and T=48 hours, it was determined that the[¹⁴C]-Compound A dosing solution was stable for at least 48 hoursprotected from light at refrigerated conditions. Degradation wasobserved at 48 hours for the [¹⁴C]-Compound A solution that was storedat room temperature and protected from light.

The final formulation for the [¹⁴C]-Compound A dosing solution wasdeveloped to deliver a single capsule containing a solution of 20 mg ofCompound A with a microtracer of [¹⁴C]-Compound A (200 nCi).

The formulation was prepared using 50:50 (v:v) EtOH:PEG 400,[¹⁴C]-Compound A, and Compound A drug substance to achieve a finalconcentration of 28.6 mg/mL. Preliminary stability data indicates thatthis formulation was stable for at least 48 hours when stored atrefrigerated conditions and protected from light.

6.5 Biological Examples

6.5.1 A Phase 1, Open-Label, Randomized, Crossover Study to Evaluate thePharmacokinetics of Compound A after A Single Oral Dose of Tablet andCapsule Formulations in Healthy Male Adult Subjects.

Certain formulations provided herein were evaluated in a Phase 1,open-label, randomized, crossover study. The study had a Screeningphase, three Treatment and Sample Collection periods, and a follow-upvisit.

Within no more than 21 days (Day −21) and no less than 2 days (Day −2)prior to the start of Period 1, subjects underwent routine screeningprocedures including physical examination, 12-lead electrocardiogram(ECG), assessment of vital signs, clinical laboratory safety tests(serum chemistry, hematology, and urinalysis), serology screen, fastingglucose levels and drug/alcohol screen.

Eligible subjects returned to the study center on Day −1 of Period 1 forbaseline assessments. During each study period, subjects were domiciledat the study center from Day −1 through Day 5. Subjects were dischargedfrom the study center on the morning of Day 5 upon satisfactory safetyreview and completion of study-related procedures.

On Day 1 of Period 1, following an overnight fast of at least 8 hours,subjects were randomized to one of the following 3 sequences to receiveTreatment A, B or C (Table 35).

TABLE 35 Treatment Sequences Period 1 Period 2 Period 3 Sequence 1 A B CSequence 2 B C A Sequence 3 C A B

In Treatment A, one 20-mg reference Compound A API-in-capsule wasadministered orally after at least 8 hour fast with 240 mL ofnon-carbonated, room temperature water. In Treatment B, one 20-mg tabletof Compound A (Tablet Formulation XIII) was administered under fastedconditions. In Treatment C, four 5-mg tablets of Compound A (TabletFormulation XIV) were administered under fasted conditions. The 20-mgtablet and four 5-mg tablets were administered orally after at least 8hour fast with 240 mL of non-carbonated, room temperature water.

The periods were separated by a washout period of at least 7 days (nomore than 10 days) from the prior dose to the next dose. In certaininstances, a longer washout is acceptable.

For each period, serial blood samples were collected before dosing (zerohour) and at 0.5, 1, 1.5, 2, 2.5, 3, 4, 6, 8, 12, 24, 48, 72, and 96hours after dosing. Plasma concentrations of Compound A were determinedfor determining PK parameters, such as AUC_(0-t), AUC_(0-∞), C_(max),T_(max), t_(1/2), CL/F, and Vz/F for Compound A. Plasma PK parameterswere calculated using non compartmental methods. Analyses of variance(ANOVA) were performed on the natural log-transformed AUC_(0-t),AUC_(0-∞), and C_(max) for Compound A. The geometric mean ratios(test/reference) and their 90% confidence intervals were alsocalculated. For T_(max), non parametric analysis was used to producemedian differences.

Blood samples to assess PD were collected at Baseline (Day −1) in Period1 for all subjects. After randomization, serial PD blood samples werecollected only in each period in which Treatment B (20 mg tabletformulation) was administered. Samples were collected prior to dosing(zero hour) and at 1.5, 3, 6, 8, 12, 24, and 48 hours afteradministration of Treatment B. The samples were used for biomarkeranalysis which involve measuring levels of pAKT (mTORC2), p4EB-P1,and/or pS6RP (mTORC1); and/or and pAKT (mTORC2) by flow cytometry usingwhole blood samples and/or other exploratory biomarkers in pre- andpost-treatment samples at different time points. The biomarker data wereused for exploration of PK-PD relationships.

Safety was monitored throughout the study. Safety evaluations includedAE reporting, physical examinations, vital sign measurements, ECGs, andclinical laboratory safety tests. Concomitant medications were assessedand recorded throughout the study from the time informed consent wasobtained until the follow-up visit.

All subjects returned to the clinic within 7 to 10 days after the lastdose in Period 3 for follow-up safety assessments. In the event that asubject discontinued prematurely from the study, every reasonable effortwas made (and documented) to ensure that all procedures and evaluationsscheduled for the follow-up visit were performed at time ofdiscontinuation or a follow-up visit was scheduled within 7 to 10 daysfrom the discontinuation day.

Results: The major PK parameters are summarized in Table 36 and Table 37(see FIG. 8 for plasma concentration-time profiles).

TABLE 36 Pharmacokinetic Parameters (Geometric Mean (Geometric CV %))Treatment A (n = 18) O-Desmethyl Treatment B Treatment C Parameter Cmpd.A metabolite (n = 17) (n = 17) T_(max)* (h) 1.5 (1-3) 3.0 (2-24) 1.5(1-2.5) 1.00 (1-3) C_(max) (ng/mL) 190 (20) 503 (24) 198 (22) 212 (29)AUC_(0-∞) (ng · h/mL) 985 (26) 11928 (23) 988 (27) 980 (30) AUC₀₋₂₄ (ng· h/mL) 934 (24) 7484 (22) 944 (26) 938 (29) Vz/F (L) 167 (28) ND 161(28) 158 (30) CL/F (L/h) 20.3 (23) ND 20.2 (27) 20.4 (30) t_(1/2) (h)5.7 (24) 14.3 (20) 5.6 (22) 5.4 (23) *T_(max) presented as median(range).

TABLE 37 90% CI of Intra- Geometric Ratio of Ratio (%) of SubjectParameter Treatment N Mean Means Means CV % AUC_(0-t) A 18 941.2 99.7 (Bvs A) 94.7-105.0 8.9 (ng · h/mL) B 17 938.5 C 17 934.7 99.3 (C vs A)94.3-104.6 AUC_(∞) A 18 985.4 99.3 (B vs A) 94.8-104.0 8.0 (ng · h/mL) B17 978.4 C 17 969.8 98.4 (C vs A) 94.0-103.1 C_(max) A 18 190.2 103.8 (Bvs A) 93.6-115.0 17.9 (ng/mL) B 17 197.4 C 17 212.3 111.6 (C vs A)100.7-123.7  Abbreviations: AUC_(∞) = area under the plasmaconcentration versus time curve from time zero to infinity; AUC_(0-t) =area under the plasma concentration versus time curve from time 0 to thelast quantifiable concentration; CI = confidence interval.

Conclusions: Compound A pharmacokinetics are comparable after singledose administration of 20 mg Compound A tablet formulations and API incapsule in healthy adult male subjects.

6.5.2 A Phase 1, Open-Label Study to Evaluate the Metabolism andExcretion of Compound A and the Effect of Food on the Pharmacokineticsof Compound A in Healthy Male Adult Subjects.

The primary objectives of this study are: to characterize thebiotransformation and excretion of Compound A following a single 20 mgoral dose of Compound A capsule containing a microtracer of[¹⁴C]-Compound A solution in healthy male subjects (Part 1) and toevaluate the effect of a high-fat meal on the pharmacokinetics (PK) ofCompound A following a single oral 20-mg dose of Compound A tablet (Part2).

The secondary objectives of this study are to evaluate the tolerabilityof Compound A after a single 20-mg oral dose of Compound A capsulecontaining a microtracer of [¹⁴C]-Compound A solution in healthy maleadult subjects (Part 1), to evaluate the effect of a high-fat meal onthe PK of the O-desmethyl metabolite of Compound A following a single20-mg oral dose of Compound A tablet (Part 2) and to evaluate thetolerability of Compound A after a single 20-mg oral dose of Compound Atablet in healthy male adult subjects (Part 2).

The primary endpoints of Part 1 are: Total [¹⁴C]-radioactivity in wholeblood, plasma, urine and feces; cumulative excretion of Total[¹⁴C]-radioactivity (as fraction of radioactive dose) in urine andfeces; total [¹⁴C]-radioactivity whole blood-to-plasma ratios;concentration of Compound A and the O-desmethyl metabolite of Compound Ain plasma, urine, and feces samples collected up to 14 times from theday prior to dosing to 8 days after dosing; and metabolitecharacterization and profiling in plasma, urine and fecal samples.Plasma PK parameters for total radioactivity, Compound A and theO-desmethyl metabolite of Compound A (e.g., C_(max), T_(max), AUC_(0-t),AUC_(∞), t_(1/2)) will be determined provided sufficient data areavailable.

The primary endpoints of Part 2 are: Plasma PK parameters (e.g.,C_(max), T_(max), AUC_(0-∞), t_(1/2)) for Compound A and the O-desmethylmetabolite of Compound A under fed and fasted conditions.

The shared secondary endpoints of Part 1 and Part 2 are: Adverse event(AE) reporting (includes serious AE [SAE] reporting); Complete physicalexaminations; Clinical laboratory safety tests; Vital sign measurements;12-lead electrocardiograms (ECGs); and Concomitant medications.

The secondary endpoint of Part 2 is: Plasma PK parameters (e.g.,C_(max), T_(max), AUC_(0-t), AUC_(∞), t_(1/2)) for the O-desmethylmetabolite of Compound A under fed and fasted conditions.

This will be a single-center, 2-part, open-label, randomized (Part 1only), 2-treatment study in healthy adult males (n=18). Within no morethan 28 days (Day−28) prior to the start of Part 1 or Part 2, subjectswill undergo routine screening procedures including physicalexamination, 12-lead electrocardiograms (ECGs), vital signs, clinicallaboratory safety tests (plasma or serum chemistry, hematology, andurinalysis), serology screen, fasting glucose levels (including HbA1C)and drug and alcohol screen.

On Day 1 of Part 1, subjects who continue to be qualified forparticipation in the study will be enrolled following an overnight fastof at least 8 hours. For Part 2 and on Day 1 of Period 1, subjects whocontinue to be qualified for participation in the study will be randomlyassigned to 1 of 2 treatment sequences (Cohort 2 or Cohort 3) andenrolled in Part 2 following an overnight fast of at least 8 hours.Subjects will be enrolled in Part 1) and Part 2) to receive Treatment Aor B in one of the following 3 cohorts:

Study Part Cohort Period 1 Period 2 Part 1 Cohort 1 (n = 6) Treatment A(fasted) NA Part 2 Cohort 2 (n = 6) Treatment B (fasted) Treatment B(fed) Cohort 3 (n = 6) Treatment B (fed) Treatment B (fasted) TreatmentA: A single 20-mg oral dose of Compound A capsule containing amicrotracer of [¹⁴C]-Compound A solution under fasted conditions.Treatment B: A single 20-mg oral dose of Compound A tablet under fastingor fed conditions.

Part 1 design: After screening, subjects (n=6) eligible forparticipation in the study will return to the study center on Day −1 forbaseline assessments. Subjects who continue to be qualified forparticipation in the study will be enrolled in the study on the morningof Day 1. Subjects will receive Treatment A after fasting overnight forat least 8 hours and will continue fasting (not consume any food) until4 hours after dosing on the morning of Day 1. Water will be allowedduring the fasting period. Subjects will be domiciled at the studycenter from Day −1 until the morning of Day 8. Subjects will bedischarged from the study center on the morning of Day 8 uponsatisfactory safety review and completion of study-related procedures.

Serial blood samples (10 mL) will be collected at predose (0 hour) andat 0.5, 1, 2, 3, 6, 12, 24, 48, 72, 96, 120, 144, and 168 hours postdose. Total [¹⁴C]-radioactivity will be determined in blood, plasma,urine and feces. Blood-to-plasma ratios will be calculated to determinepartitioning for total [14C]-radioactivity. Urine samples will becollected at predose (within 2 hours prior to dose administration) andat the following post dose collection intervals: 0 to 6, 6 to 12, 12 to24, 24 to 48, 48 to 72, 72 to 96, 96 to 120, 120 to 144, 144 to 168hours. Total urine volume collected in each interval will be recordedfor determination of the fraction of dose excreted in urine. All fecalsamples will be collected daily from Day −1 through Day 8 and weight ofdaily fecal collections will be pooled and recorded.

Part 2 design: Part 2 will be a 2-period crossover study; in Period 1,subjects (n=12) will be randomized to receive either an oral 20 mg doseof Compound A tablet (Treatment B) under fed (n=6) or fasted (n=6)conditions. In Period 2, subjects will receive Treatment B underconverse conditions based on treatment assignment in Period 1. Afterscreening, subjects (n=12) eligible for participation in the study willreturn to the study center on Day −1 for baseline assessments. Subjectswho continue to be qualified for participation in the study will berandomized and enrolled in the study on the morning of Day 1. Subjects(n=6) will be enrolled and randomized to receive Treatment B under fedor fasted conditions on the morning of Day 1 after fasting for at least8 hours. Fed subjects will be served a standard high fat meal breakfast,or its equivalent, that must be consumed within 30 minutes from serving.Dosing must occur 30 minutes (+5 minutes) after serving a subjectbreakfast. All subjects (fed and fasted) will fast (not to consume anyfood) until 4 hours post dose. Water will be allowed during the fastingperiod. Subjects will be domiciled at the study center from Day −1 untilthe morning of Day 5 of each period. Subjects will be discharged fromthe study center on the morning of Day 5 upon satisfactory safety reviewand completion of study-related procedures. Safety and tolerability datawill be monitored and collected following each dosing period. Periods 1and 2 will be separated by a washout period of at least 7 days (no morethan 10 days) from prior dose to the next dose. In certain instances, alonger washout may be acceptable if previously agreed to.

Serial blood samples (10 mL) will be collected at predose (0 hour) andat 0.5, 1, 2, 3, 6, 12, 24, 48, 72 and 96 hours post dose for thedetermination of plasma concentrations of Compound A and the O-desmethylmetabolite of Compound A. Safety will be monitored throughout the study;safety evaluations will include AE reporting, physical examinations,vital sign measurements, ECG, and clinical laboratory safety tests.Concomitant medications will be assessed and recorded throughout thestudy as well. In addition, during the subjects' stay-in-the clinic(i.e., confinement period), fasting plasma glucose levels will bemonitored as part of the clinical laboratory safety tests. For Parts 1and 2, all subjects will return to the clinic within 7 to 10 days afterthe last dose for follow up safety assessments. In the event that asubject discontinues prematurely from the study, every reasonable effortshould be made (and documented) to ensure that all procedures andevaluations scheduled for the follow-up visit are performed at the timeof discontinuation or a follow-up visit should be scheduled within 7 to10 days from the discontinuation day.

Part 1 dosing: Subjects will fast overnight for at least 8 hours priorto Compound A administration. On the morning of Day 1, each subject willbe dosed under fasting conditions with a single 20-mg oral dose ofCompound A capsule containing microtracer of [¹⁴C]-Compound A in anethanol/polyethylene glycol solution. The exact specific activity,chemical purity and radiochemical purity will be determined prior todosing. After dosing, subjects will continue to fast until 4 hours afterdosing; thereafter, they will be served standard meals and snacks.Dosing time will be recorded in the source documents and CRF. Dosinginstruction and calculation for actual dose administered to each subjectwill be provided at or before study initiation. The actual dose of the[¹⁴C]-Compound A microtracer administered to each subject will becalculated based on the measured radioactivity concentration (dpm/g) ofthe solution in the capsule.

Part 2 dosing: In Part 2, subjects will fast overnight for at least 8hours prior to Compound A administration. On the morning of Day 1, eachsubject will receive a 20-mg tablet of Compound A orally. Subjectsrandomized to receive Compound A under fed conditions will be a served astandard high fat meal (breakfast).

The standard high fat meal or its equivalent must be consumed within 30minutes of serving. Dosing must occur 30 minutes (+5 minutes) afterserving the meal. The tablet will be administered with approximately 240mL of non-carbonated, room temperature water. After dosing, subjectswill continue to fast until 4 hours after dosing.

Subjects enrolled in the study will spend a total of approximately 8weeks on the study.

Subjects must satisfy all of the following inclusion criteria to beeligible for enrollment into the study: 1. Must understand andvoluntarily sign a written ICD prior to any study-related proceduresbeing performed and be able to adhere to restrictions and examinationschedules; 2. Must be able to communicate with the investigator andclinical staff and to understand and comply with the requirements of thestudy; 3. Must be a male 18 to 55 years of age (inclusive) at the timeof signing, with a BMI (weight (kg)/(height (m²)) between 18 and 33kg/m² (inclusive) and weight between 60 and 100 kg (132 to 220 lbs;inclusive); 4. Must be healthy (at Screening and Day −1) as determinedby the investigator on the basis of medical history, physicalexamination, clinical laboratory safety test results, vital signs, and12 lead ECG (Vital signs (systolic and diastolic blood pressure, pulserate, and oral body temperature) will be assessed in the supine positionafter the subject has rested for at least 5 minutes, Subject must beafebrile (febrile is defined as ≥38.5° C. or 101.3 Fahrenheit), Systolicblood pressure in the range of 90 to 140 mmHg, diastolic blood pressurein the range of 60 to 90 mmHg, and pulse rate in the range of 45 to 100bpm, Screening fasting plasma glucose value within the normal limits ofthe institution and HbA1C<6%); 5. Subjects (with or without vasectomy)must agree to use barrier contraception (i.e., latex condom or anynon-latex condom not made out of natural (animal) membrane (e.g.,polyurethane)) and one other method (e.g., spermicide) when engaging insexual activity with woman of child-bearing potential during studyconduct, and for 90 days after the last dose of study medication; and 6.Must agree to refrain from donating blood or plasma (other than for thisstudy) while participating in this study and for at least 28 days afterthe last dose of study drug.

The presence of any of the following will exclude a subject fromenrollment into the study: 1. Recent history (i.e., within 3 years) ofany clinically significant neurological, gastrointestinal, hepatic,renal, respiratory, cardiovascular, metabolic, endocrine, hematological,dermatological, psychological, or other major disorders; 2. Anycondition, including the presence of laboratory abnormalities, whichplaces the subject at unacceptable risk if he were to participate in thestudy, or confounds the ability to interpret data from the study; 3. Useof any prescribed systemic or topical medication within 30 days of thefirst dose; 4. Use of any non-prescribed systemic or topical medication(including herbal medicines) within 7 days of the first doseadministration (with the exception of vitamin/mineral supplements); 5.Subject used any metabolic enzyme inhibitors or inducers (i.e., CYP3Ainducers and inhibitors or St. John's Wort) within 30 days of the firstdose administration; 6. Presence of any surgical or medical conditionspossibly affecting drug absorption, distribution, metabolism, andexcretion, or plans to have elective or medical procedures during theconduct of the trial; 7. Exposure to an investigational drug (newchemical entity) within 90 days prior to the first dose administration;8. Donation of blood or plasma within 60 days prior to the first doseadministration; 9. History of multiple (i.e., 2 or more) drug allergies;10. Any clinical significant allergic disease (excluding non-active hayfever), excluding nonactive seasonal allergies and childhood asthmacleared for at least 3 years; 11. History of drug abuse within 2 yearsprior to first dosing, or positive urine drug screening test due toillicit drugs; 12. History of alcohol abuse within 2 years prior todosing, or positive alcohol screen; 13. Smokes more than 10 cigarettes,or consumes the equivalent in tobacco, per day; 14. Known to have, ortests positive for, active or chronic hepatitis B or hepatitis C, or HIVantibodies; 15. Received vaccination (excluding seasonal fluvaccination) within 90 days of the study drug administration; or 16. ForPart 1 only: Prior exposure to radioactive investigational drugs within6 months prior to check in, and prior exposure to work-related,diagnostic or therapeutic radiation within 12 months prior to check in.

Inclusion/exclusion criteria will be assessed at screening. Subjecteligibility will be confirmed again on the admission day (Day −1) of thefirst period and/or prior to randomization on Day 1 by physicalexamination, drug screen, clinical laboratory safety tests vital signsand ECGs.

Preliminary Results: 11/12 enrolled subjects completed Part 2. Theresults are set forth in Table 38, below.

TABLE 38 Geometric Mean CV %) Pharmacokinetic Parameters After Single20-mg Oral Dose Fasted (n = 11) Fed (n = 11) O-Desmethyl O-DesmethylParameter Cmpd. A metabolite Cmpd. A metabolite T_(max)* (h) 1.00 (1-2)3.00 (1-3) 3.00 (1-3) 6.00 (3-12) C_(max) (ng/mL) 182 (24) 425 (23) 156(27) 364 (28) AUC_(inf) (ng*h/mL) 1005 (38) 9834 (38) 1195 (38) 10131(35) AUC₀₋₂₄ (ng*h/mL) 955 (35) 6401 (30) 1131 (34) 6271 (29) Vz/F (L)151 (28) 34.7 (28) 125 (20) 42.6 (30) CL/F (L/h) 19.9 (38) 2.0 (38) 16.7(38) 2.0 (36) t_(1/2) (h) 5.3 (33) 14.8 (25) 5.2 (27) 14.9 (29) *T_(max)presented as median (range).

Conclusions: After administration of Compound A with a high fat meal tohealthy adult males, there is an approximate 17% decrease in Compound AC_(max) and an approximate 20% increase in overall exposure (AUC_(inf)).There is also a 2 hour delay in T_(max). After administration ofCompound A with a high fat meal to healthy adult males, there is anapproximate 17% decrease in O-desmethyl metabolite C_(max) and anapproximate 3% increase in overall exposure (AUC_(inf)). There is also a3 hour delay in T_(max).

The embodiments disclosed herein are not to be limited in scope by thespecific embodiments disclosed in the examples which are intended asillustrations of a few aspects of the disclosed embodiments and anyembodiments that are functionally equivalent are encompassed by thepresent disclosure. Indeed, various modifications of the embodimentsdisclosed herein are in addition to those shown and described hereinwill become apparent to those skilled in the art and are intended tofall within the scope of the appended claims.

A number of references have been cited, the disclosures of which areincorporated herein by reference in their entirety.

What is claimed is:
 1. A crystal form of the compound of formula (I),7-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1-((trans)-4-methoxycyclohexyl)-3,4-dihydropyrazino[2,3-b]pyrazin-2(1H)-one,or a salt thereof:

which has an X-ray powder diffraction pattern comprising peaks atapproximately 7.46, 13.70 and 23.06° 2θ.
 2. The crystal form of claim 1,which has an X-ray powder diffraction pattern further comprising peaksat approximately 8.94, 18.22 and 18.78° 2θ.
 3. The crystal form of claim1, which has a thermogravimetric analysis thermogram comprising a totalmass loss of approximately 11.0% of the total mass of the crystal formwhen heated from about 25° C. to about 300° C.
 4. The crystal form ofclaim 1, which has a single differential thermal analysis thermogramcomprising an endotherm between about 90° C. and about 125° C. with amaximum at approximately 106-110° C. when heated from about 25° C. toabout 300° C.
 5. The crystal form of claim 4, wherein the singledifferential thermal analysis thermogram further comprises an endothermwith a maximum at approximately 193° C.
 6. The crystal form of claim 1,which is p-xylene solvated.
 7. The crystal form of claim 6, whichcomprises 0.5 molar equivalents of p-xylene.
 8. The crystal form ofclaim 1, which is substantially pure.